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KingFisher mL - User Manual

1. Thermo Scientific KingFisher mL User Manual Rev 1 3 Table of Contents Thermo Scientific KingFisher mL Cat no 5400050 User Manual Rev 1 3 August 2007 Cat no 1508260 Contents 1 Safety Symbols and Markings rrerrnnvnnnnnnnvnvnnvrnnvnnennnnvnvensnnvnversnnvnnensnnvnnnssnnnnnennnnnnnnersnnnnnnnnnen 6 2 About the User Manual rernnnvovnnnnnvvvnnnnnvevnnnnnvvnnnnnnvvnnnnnnnvnnnnnnnvnnnnnnnvnnnnnnnnnnnnnnsvnnnnnnsvvnnnnnnnnvennn 7 3 Introduction to the KingFisher mL rrrunnnnnnnnnvnnnnnnnnnnnnnvnvnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 8 4 Functional DeScriptioni t ci 2c c5 feccectscsscsatdiegscxdsdusncnuenencecarsssneondctusacussdescenecanessuensunetadsucerceddswacedecens 9 5 Installation AIC EDD 14 6 Routine Operation iz 5 1 nein E Aaa T ERet D Ri ARI EN E AAAA nae 18 7 Maintenance RII EET 22 8 Technical Specifications 2 5 gnus idad 25 9 Troubleshooting Guide snrnnnnnvnvnnnnnnnvnnnnnnnnvnnnnnnnvnnnnnnnvnnsnnnnennnnnnnvnnnnnnnvnnennneneernnennennneevennnnesn 28 10 Warranty Certificate iii iia 33 11 Ordering Information niei eere ee An rt cas Soda dan RII E RE NR AINE KRARASINRRRRRDMRR RR RAS CR dana 34 12 Glossary and AbbreviatiONS coonocccnnnnncccononccccnnonccnoncnnoncncnnnnnnncncnnn en nnne nn nn nnne nn en nnne nnne nnne nnn 35 13 ub ELEC E TA 37 14 Table Of Figures edem ts 40 15 Appendix A Application Protocols for KingFisher mL r
2. Cat no Serial no PURCHASED BY Ge Company Institute Distributor Department Address Address Tel Tel Fax Date of delivery Internet home page Date of purchase Your research area Dr Mr L1 Mrs O Ms O Job title Position Surname block capitals First name block capitals Internet e mail address Excellent Above As Below Comments expectations expected expectations Reagent kit Instructions O L1 O L1 Instrument User manual O O L1 L1 Operational reliability O O L1 O Design O O O O Ease of use Li O L1 L1 Operational costs o O L1 a Customer support O O O L1 Further instrument system developments desired Further applications desired Where did you first learn about the product Would you like to receive information about other Thermo Scientific products Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 45 Appendix D Addresses 18 Appendix D Addresses For the latest information on products and services visit our websites at http www thermo com http www thermo com kingfisher Distributed by Manufactured by Thermo Fisher Scientific Oy P O Box 100 F 01621 Vantaa Finland Tel 4 358 9 329 100 Fax 358 9 3291 0415 www thermo com KingFisher Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 46 Appendix E Supplementary Application Protocols 19 Appendix E Supplementary Application Protocols Thermo Scientific KingFisher m
3. a a a non an nn cnn aran nn rra arar rra 22 How 10 p ck for SERVICE ii A E A 24 31 How to release the transport lock oooococcccnnnnccoconnnnncnnnannnnnnnnccnnnnnnnnnnn eae ee eee ea ta rre 15 How to setup the instrument before you put it into OperatiON oonoccccnnnoccccnanonnnnnnanoncnn nano ano nanaranonnnno 15 a ohn AoE aUo LOATA a ASSA N EEA Ea EAA A a daa aed a cR 21 HOWt SWIteh OM ERE PERE di T 18 20 How to troubleshoot the instrument essen nnne nnns kann disais seda anna sana na 29 How to Unpack etre reu e Ee KATT FOIE ain RETE ir OI rM ins 14 How to use the keypad and display ooccooconccocccccnnnnonanononcnnnoncnnnnnnnnn corn c nn eene nennen rra 18 How to work with a magnetic rod cccccccccececcececececececececececececesececesecauaeaeauaeaeaeaeaeaeaeaeaeaeauaeaauansaaaensaeass 12 IncubatioD i2 et ee A ee thea ette vto eta tede EO EE et da re Ted etus 8 12 13 25 42 Sai A A E 7 14 15 24 26 Instrument layo te sinana a ria 9 Intended US aunn a EC the bl eme EE La en Ede e Ee bem s n sjette sein Ea bd e cas 8 Inverse magnetic particle processing occononococcnnnoncnonnnonencnnnnnnnnnnnn nn nn ae nnn nana nennen nnne nennt nnn enn 11 12 K Keypad and display e ete Fluenes 8 11 18 44 M CN 18 25 41 Keywords for web pages nis rr a RUE e PAPE MA XXE anio 35 dec aa 8 19 33 45 L Liguidierystal displays ih rer rrt dick c
4. e Defects caused by fire lightning flood earthquake explosion sabotage war riot or any other occurrence of the type listed above e Refurbished products that are subject to different warranty conditions THIS WARRANTY IS IN LIEU OF ALL OTHER EXPRESSED OR IMPLIED WARRANTIES INCLUDING BUT NOT LIMITED TO ANY IMPLIED WARRANTIES OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE The seller is not liable for any loss or damage arising out of or in connection with the use of the product or other indirect damages These warranty terms and conditions can be obtained from your local Thermo Fisher Scientific dealer 10 1 Warranty limitations e Consumables are not included in the warranty e The warranty does not extend beyond the expiration date of the reagent kit Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 33 11 Ordering Information Code 5400050 Instrument System KingFisher mL 100 240 V 11 1 List of accessories Code 1508260 97 002 111 97 002 121 97 002 131 97 002 141 4540000 9404190 Item KingFisher mL User Manual KingFisher mL tip comb KingFisher mL tube Chapter 11 Ordering Information Quantity 1 800 pcs bag 900 pcs bag 15 x 60 pcs KingFisher mL Combi 60 tubes and tip combs for 60 samples box KingFisher mL Combi 240 tubes and tip combs for 240 samples box FP Stepper 10 ul 5 ml Finntip 2 5 ml 11 2 List of spare parts Code 11273100 24073180
5. Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 36 Chapter 13 Index 13 Index A Abbreviatlons Li il 35 About the User Manuals copo iria itte itn Ea e a Een b EF aan cidad dad 7 AGCCOSSOLT OGS cci ite studie dernest A ga ecc eda aver Fed e rea a 14 34 Addtesses diras 46 Advantages of processing magnetic Particles scccccccccscsssssscsececececsssessneeceseesecesesseaeeeeesesssessaaeeness 8 Application OA FO seated Te shad i na AA s Eee rat ide 5 6 7 36 41 45 47 Applicat o o Hojto eo ARR PERO 7 41 Cell or protein p r aT O E a e nennen Aa nnnm nennen nnnn nasa ares nest sa a aaa Randen sas anna a 41 KingFisher mL total RNA purification sssesessseseeeseeee eene eene nnne nnn nnnnna naar nass nenas nsa nass sanas 42 B Brief User s Gulde ie d rc m exa rer e an a D ava Cur CX ER E o PR NDA EE rae eden 44 C Gable C 17 23 34 Certificate of DECONTAMINATION sees ana nan nn cnn narran anna rr rra 24 31 32 GROCKS cans EEE EE E aguas vena duanibdaaecavbua came etaweadsband came veutbacveiaanes 14 17 29 30 31 Clean da ha aden EE AT 15 22 23 30 magnetierodsuastsv ER 22 processing chamber ss4 Ae 22 Concentration E ta lata a 12 13 43 D DECOMAMIN AMO A age i RE V FINA E iia 24 31 32 DIMENSIONS iia a aaa du RR da E o 2b DirectiV8S icis iiti rhe retener here ZG E ia pi tee lb Ea bd exea bl va atm bie ar Teen Saabs SER CAPE aga tn Xa Fr n 27 DIS
6. 2305290 5186980 Item Quantity Tube strip tray Tube rack Microtube tray Serial cable F9 F25 for RS 232C port KingFisher Software version 1 0 a 11 3 List of recommended spare parts Code 1091910 1092250 2004460 2004470 2004480 21040811 21040820 21040850 24070740 24072550 ltem 1 2 unit s year Tooth belt T2 5 330 Tooth belt T2 5 265 PCB PSP 01 PCB CSU 02 1 PCB CSU 01 Power supply 1 H motor assembly M motor assembly Free wheel assembly 1 Magnetic plate assembly Contact the Thermo Fisher Scientific service department 1 100 pcs box 10 units year SU gt gt N gt Naao Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 34 12 Glossary and Abbreviations Chapter 12 Glossary and Abbreviations Magnetic rod The rods which are magnetic and collect magnetic particles The rods do not collect the particles on their own the magnetic rods must always be protected by a tip comb Tip comb Protects the magnetic rod A disposable tip comb always has to be fitted into the tip comb holder slot during processing Tube strip The disposable tubes where all the reagents and samples are located and where the processing takes place Tube strip tray A detachable tube strip holder or tube rack that can be placed on the Tube rack laboratory bench to facilitate pipetting Fig 6 3 and Fig 6 5 12 1 Keywords for web pages affinity separation amplif
7. combs KingFisher mi Fig 4 5 KingFisher mL magnetic particle processor Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 11 Chapter 4 Functional Description 4 3 Principle of magnetic particle processing Magnetic rod ld 7 n j P ld ld 4 P Tip pl Li i 1 4 Id Li ld j y p J y 4 eii 4 Mi ld 4 ar Transfer Wu Jm dr 4 ld 4 4 la i C f il a J gt y 4 4 y y Or 7 ir I T A y Jj E r y A f Mi wot Y 4 Y n EA d f Y ot pi 4 Y f Y P 1 Y gl AA f A 4 Pi di 4 f iV 4 f y 4 s r 1 y t a JA z A Y Y a 4 7 y ry 44 n 4 4 LY 4 Y Magnetic particle Collection of Magnetic particles Release of Ready suspension before magnetic particles on the tip surface magnetic particles suspension collection Fig 4 6 Inverse magnetic particle processing 4 4 Working with a magnetic rod Working with magnetic particles can be divided into five separate processes collecting particles releasing particles washing particles incubation concentration 4 4 1 Collecting magnetic particles During the collection of the magnetic particles the magnetic rod bar is fully inside the tip The magnetic rods together with the tip comb s move slowly up and down in the tubes and the magnetic particles are collect
8. 1 3 How to clean the processing chamber If required clean the processing chamber using a soft cloth or tissue paper soaked in a mild detergent solution SDS soap solution or alcohol Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 22 Chapter 7 Maintenance 7 2 Disposal of materials Refer to local regulations for the disposal of infectious material The samples can be potentially infectious Dispose of all used tube strips disposable AN gloves syringes disposable tips etc as biohazardous waste 7 3 Decontamination procedure If there is any risk of contamination with biohazardous material the procedure recommended below or some other corresponding decontamination procedure must be performed We strongly recommend that the complete decontamination procedure is performed before relocating the instrument from one laboratory to another Decontamination is not required for the proper functioning of the instrument Example of decontaminants Ethanol 70 Virkon solution 1 3 Glutaraldehyde solution 4 Chloramine T Microcide SQ 1 64 Always use disposable gloves and protective clothing and operate in a well ventilated area 1 Prepare the decontaminant for example 1 3 Virkon solution or 200 ml 4 glutaraldehyde solution or another agent recommended by your safety officer 2 Empty the tube strip tray 3 Switch OFF the power and disconnect the mains supply cable Fig 5 3 4 Disinf
9. Signature Name block capitals The signature of a Radiation Safety Officer is also required when the unit has been used with radioactive materials This unit is certified by the undersigned to be free of radioactive contamination Date and place Signature Name block capitals res PHOTOCOPIABLE Please include decontaminating solution used Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 32 Chapter 10 Warranty Certificate 10 Warranty Certificate Thermo Fisher Scientific Microplate Instrumentation Business products are fully guaranteed against defective parts and materials including defects caused by poor workmanship for a period of one year from the date of delivery Thermo Fisher Scientific will repair or replace defective parts or materials during the term of warranty at no extra charge for materials and labor provided that the products were used and maintained in accordance with Thermo Fisher Scientific s instructions The warranty is invalid if products have been misused or abused For the warranty to be effective the product must have been purchased either directly from Thermo Fisher Scientific or from an authorized Thermo Fisher Scientific distributor The guarantee is not transferable to a third party without prior written approval from Thermo Fisher Scientific This guarantee is subject to the following exclusions e Any defects caused by normal wear and tear
10. low pressure compressed air or a cloth dampened with water or a mild detergent when necessary Although the KingFisher mL is constructed from high quality materials you must immediately wipe away spilt saline solutions solvents acids or alkaline solutions from outer surfaces to prevent damage Painted surfaces can be cleaned with most laboratory detergents Dilute the cleaning agent as recommended by the manufacturer Do not expose painted surfaces to concentrated acids or alcohols for prolonged periods of time as damage may occur Clean the display areas with a mild laboratory detergent The keypad has a wipe clean surface Plastic covers and surfaces can be cleaned with a mild laboratory detergent or alcohol If any surfaces have been contaminated with biohazardous material a mild sterilizing A solution should be used Do not autoclave any part of this instrument except for the tube strip tray 7 1 1 How to clean the tube strip tray Keep the tube strip tray surface clean to avoid dust and dirt entering into the instrument Clean the tray surface at least once a week using a soft cloth or tissue paper soaked in a mild detergent solution SDS soap solution or alcohol If you have spilt infectious agents on the tray you can autoclave the tray 7 1 2 How to clean the magnetic rods If required wipe the magnetic rods using a soft cloth or tissue paper soaked in a mild detergent solution SDS soap solution or alcohol 7
11. mL system do A1 Thermo Fisher Scientific now offers a complete KingFisher mL system the magnetic particle processor for the purification and processing of DNA RNA cells and proteins The processor handles particles automatically according to the preloaded purification protocols For more information on these applications contact Thermo Fisher Scientific Oy Q2 Do I need a PC to control the processor A2 Normally the protocol s is are preloaded by Thermo Fisher Scientific and no PC is needed for routine use KingFisher Software can be used to create new protocols or modify existing protocols if needed PC modified protocols are then downloaded into the processor for routine use Q3 Will the magnet get weaker If so how long can they be used Can they be replaced A3 The KingFisher mL magnets are made of material that is very stable The magnetic field will not get weaker Magnets can be replaced if needed Usually there is no need to replace them but it can be done for example if they are mechanically damaged during runs Changing the magnets can be carried out by local technical personnel 04 How strong are the magnets Can they for example disturb some sensitive equipment A4 The magnetic field is situated just around the magnetic tips so it is very local Therefore the magnetic field should not cause any problems to surrounding equipment However if you touch your floppy disk with the magnetic rod you will lose
12. tieetosstes 21 16 7 KingFisher mL keypad arin ipti i pide ee age te eves 44 Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 40 Appendix A Application Protocols for KingFisher mL 15 Appendix A Application Protocols for KingFisher mL A 1 Example of a cell or protein purification protocol using KingFisher mL Cell separation and protein purification based on paramagnetic particles can be automated with the KingFisher mL magnetic particle processor Generate a protocol with Thermo Scientific KingFisher Software for cell or protein isolation called General_protocol following the instructions below Load the protocol into the internal memory of the instrument Pipette the reagents into the Thermo Scientific KingFisher mL tube strips After you have filled the KingFisher mL tube strips insert the tray into the KingFisher mL processor and choose the correct program by using the arrow keys The name of the KingFisher mL protocol is General_Protocol2 Start the protocol by pressing the START button Purified cells or proteins are released into a 200 ul volume into the last tube 5 The description of the KingFisher mL protocol is as follows Program 1 The sample is mixed with particles in tube 1 2 The sample is incubated with particles for 3 minutes in tube 1 3 The particles are washed in washing buffer in tube 2 4 The particles are washed in washing buffer in tube 3 5 The particles are washed in washing buff
13. 2 Safety SPECIFICATIONS 5 1 rri ebrii d lar 26 8 3 In conformity with the requirements ccoooocccconoccnncnaninnncnaninnncnanonnnccnnnannrcrnnn nnne nnns 27 9 Troubleshooting G ide 5 eie erede cesses en keine e eR Rn Ra FR Dd a nere oa Rea den CR Rae nn Urs 28 9 1 Error messages and warnings ccccconconononccnnnnnnnnnnnnnnnnncnnn nn nana 28 92 Froubleshooting guide tidad ta td td 29 9 3 Frequently asked questions FAQ about KingFisher mL processing esss 30 9 4 Service request Protocol knivingen enn Fare ena ie 31 9 4 1 Decontamination ei a eite hee aere ann 31 9 4 1 1 Certificate of Decontamination cccccccnnnnnonnnnnnnnnnnnnanccnnnnnnnnnnnnnnnnnn nena rn n rr nnne nnn nr rr rre 32 10 Warranty Certificate soiree ccce e dede aeo Yan eco Rupe Naci 33 10 1 Warranty litnirtati ns x cete eco eet A e Een bee cda 33 11 Ordering Information iie coca Sided acca ers sce necp eva Souci sunsdedevissvacisedsssivasessiniscusescenuaies 34 11 1 List Of ACCESS ONES Nai A Alas 34 11 2 Listof sp re Pai A alse AAA A Aa 34 11 3 List of recommended spare parts sess sese 34 12 Glossary and Abbreviations ccieeeicie eese rain enar neam a Re eran Dk Enea ae nanen ennnen ennenen E uada 35 12 1 Keywords for web pag68S custe edited tne ee cic ir iter d ree C eh v re e 35 12 2 Lteratufeuss sata 36 13 A erten eden 37 14 EPISC lc 40 15 Appendix A Applicat
14. CLAIM GR z 5 hace oret cutie ene a roe ee Pelea eee bg ead vase reed ma ete ar CM ean EE 5 Disinfection prOGCedure iier cion eos tcu a dag vae ha nae Ra Aa eE EA ATE A EAE dido 21 23 24 31 32 IBIQ 18 Disposal of instr rment cte er inh re Hr ee er Hd Ee Pr EY FERE Fe Re Eee Re E eR wears 24 Disposal of Materials i erede A ais 23 DNA TORRENT ta as 30 35 36 E Environmental requirements circa de drit cte id ed put ea eek eva 15 Etrordmessatge 5 cire tr metre ainda 7 28 F or 30 Feedback EOEEDcs uii coca f reda vo dicen 14 45 Frequently asked questions coc remet einer a eds a aaa E NNA aa gi 30 G Genomic DNA x ni rn em Mener Sen i e DE T c ER 35 GOSS mr A ES 35 Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 37 Chapter 13 Index H How to clean the magnetic TOdS ooocccccnnnnnonononcnnncnnnnnnnnanennnn cnn nana nene 22 How to clean the processing chamber sssssessssssssssesseseeen nennen eren nnne tenenti nnns 22 How to clean the tube strip tray earannan ieta aaan aariaa a ane iraka aaan a atama Ai eak aa aaaea 22 How to disinfect the instrument veevvvrannnnnnnnnvnvrnnnnnnnnennnrenvnnnsneneerennnnnnsnesenvevrnnnnnnsseneerennnnnne 21 23 31 32 How to ensure startup iii a it it ads 17 19 How to install the KingFisher ML cccccccccccecececececececececececesesecececaeaeauauaeaeaeaeaeauaeaeaeaeaeaeaesauauanananenss 14 26 How to maintain the instrument
15. E REID LUTTER as 24 lee o o MEC etree crrerrre cree reer rere 24 31 request PrOTOCOl con ER UT ET am ERU ERR rr Eni i 31 Shutdown 255 1 iiir drea A e Deae dre tees cleans a deve Sie Pd dva revu Ped anita wid ct n eta rv Pt du dua d 21 Silica coated magnetic particles esses sees 35 Specifications sars nen AA A A ad dne TNR 7 25 26 general seinare REI OPI Ge ene 25 performance ico ee A di 25 CEN a I be en 26 STAR ina m 17 18 20 Streptavidin coated magnetic particles rrrrranarvernanavrvrnannrrernenarrenrsnnnrrenssnarrenssnarvenssnnrveessnarveessnrvesennn 35 SVIMDOIS pet 6 T Table of Figures ioni icai rh E re EE dd Ba dee UE m ERR E d Re nO En EE E Rs 40 MICA AAA Atti eee 8 10 11 12 13 19 20 21 25 29 30 34 35 44 MEA Oi A a A RR tnees 14 15 16 24 25 da MATE E E E E ann ete tne nse et deddap lending ets deel en ae 14 oe PE A ies 15 16 24 Package A A eda ta ie eee 14 24 TROUBIES OOO IM A Sua aeavatays ides 7 28 29 31 euro 29 TUBE SHIPS anna tenne eien adr ii 8 11 19 20 21 23 25 35 41 42 43 44 Tube strip TRAY ii ii vad aid 10 19 20 21 22 23 25 34 35 U Unpack 5 m gU eere EOD EE lata 14 User manual ee ccicitetuec a e nde er Uic wx Porc e ee racer 7 14 34 45 V MI M nLEE n 9 10 OA NRI ITI TITEL C 9 A O AT 10 isi bem NT 10 VOTE iaa 15 17 18 25 27 SP A O 17 18 W WAF
16. L User Manual Rev 1 3 Cat no 1508260 47 NOTES Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 48 NOTES Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 49 NOTES Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 50 Thermo Fisher Scientific Oy Microplate Instrumentation Ratastie 2 P O Box 100 FI 01621 Vantaa Finland www thermo com 1508260
17. NIN OS wont 15 16 17 28 WEN Vd 7 14 33 Wed eve ASA Daa ads 15 25 43 Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 39 Chapter 14 Table of Figures 14 Table of Figures Fig Fig Fig Fig Fig Fig Fig Fig Fig Fig Fig Fig Fig Fig Fig Fig Fig Fig 3 1 KingFisher mL magnetic particle processor esee 8 4 1 KingFisher mL front view with lids closed ssssssssssssss seen 9 4 2 KingFisher mL front view with lids open nennen nsns 9 43 KngFisliermL rear VIEW si RT ER e e Ru HT HEU e Y e e Re ES 10 44 KngFisher mbLanternal VIGW ioi PPM EP HE HEAT EHE ERE aa 10 4 5 KingFisher mL magnetic particle processor sessi nn 11 4 6 Inverse magnetic particle processing cocinar 12 4 7 A concentration step during magnetic particle processing 13 5 1 KingFisher mL transport lock tag present before release of the locks sssssssssssssss 16 5 2 Location of the transport lock screws and warning tag 16 5 3 CONNECTING NE INGINS SUP PIV Cable ea o eie e te p dd es 17 6 1 KingFisher mL keypad a RR ia 18 6 2 LOGGING INC tube strip tray si rt OE E CEDERE DEEP RE at red 19 6 3 Positioning of the tube strip tray and insertion of the tip comb s sse 19 6 4 Tip comb specially designed for the KingFisher mL sss nnns 20 6 5 Tube strip tray containing tubes specially designed for the KingFisher mL 20 6 6 Microtube KingFisher mb doubletray s i tete ted bei dte tsp
18. T mum ve Fig 6 2 Loading the tube strip tray e A tube strip contains one sample and the reagents for a single processing The user can select the position of the tube strip s on the tube strip tray The user must keep track of the sample order However the orientation of the tube strip is fixed The tube strips have to be positioned so that the slip ends face left Fig 6 3 and Fig 6 5 Note that the order in the tube strip of the sample and reagents is determined by the kit used Load only maximum fifteen special tube strips at a time into the tray for processing Fig 6 5 Slide the tray fully back in Thermo Labsystems KingFisher mL A START mem Tube strip tray with tubes Fig 6 3 Positioning of the tube strip tray and insertion of the tip comb s Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 19 Chapter 6 Routine Operation e The tip comb s always has have to be in its their slot s and fully inserted Fig 6 3 The instrument also functions with either one tube strip or one to fifteen tube strips depending on the amount of samples At least one tip comb Fig 6 4 always has to be inserted into one of the narrow slots meant for the tip comb Fig 6 4 Tip comb specially designed for the KingFisher mL e Close the front lid if you want to The front lid protects the instrument against environmental contamination Note The front lid and or top lid can be left open if des
19. all your data Generally a normal PC screen may also detect the magnetic field with a minor distortion of the image Q5 What should do if forget to insert the tip combs into the tip comb holder and the magnetic particles stick to the magnetic rods Ab Wipe the magnetic rods with a soft cloth or tissue paper soaked in a mild detergent solution soap solution or alcohol Q6 What if the magnetic particles remained in the sample well A6 If the starting material is too viscose the magnetic rods will not be able to collect the particles Dilute the sample and check that the sample is properly homogenized lysed Q7 What if the magnetic particles are attached to the tip combs A7 This happens sometimes but it will not affect the yield because the sample has been released from the particles Q8 Are the volumes of reagents in each well critical A8 We strongly recommend that you keep the given volumes within certain limits to avoid failure in performance of the chemical reactions and the processor Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 30 Chapter 9 Troubleshooting Guide 9 4 Service request protocol If the KingFisher mL requires service contact your local Thermo Fisher Scientific representative or the Thermo Fisher Scientific service department Do not under any circumstances send the instrument for service without any prior contact It is imperative to indicate the fault and nature of th
20. any time to incorporate technological developments This manual is subject to change without prior notice as part of a continuous product development Although this manual has been prepared with every precaution to ensure accuracy Thermo Fisher Scientific assumes no liability for any errors or omissions nor for any damages resulting from the application or use of this information This manual supersedes all previous editions No liability for consequential damages Thermo Fisher Scientific shall not be liable for any damages whatsoever arising out of the use or inability to use this product Contact information Thermo Fisher Scientific Oy P O Box 100 FI 01621 Vantaa Finland Tel 358 9 329 100 Fax 358 9 3291 0415 www thermo com Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 5 Chapter 1 Safety Symbols and Markings 1 Safety Symbols and Markings These symbols are intended to draw your attention to particularly important information and alert you to the presence of hazards as indicated SAFETY SYMBOLS USED IN THE KINGFISHER mL Power ON Power OFF WEEE symbol This product is required to comply with the European Union s Waste Electrical amp Electronic Equipment WEEE Directive 2002 96 EC x O WARNING MARKINGS USED IN THE DOCUMENTATION Caution risk of electric shock Caution biohazard risk Caution risk of personal injury to the operator or a safety hazard to the surrounding ar
21. cal prerequisites Place the instrument on a normal laboratory bench The net weight of the unit is approx 10 kg 23 Ibs The instrument operates at voltages of 100 240 Vac and a frequency range of 50 60 Hz 5 2 Setups before you put the instrument into operation 5 2 1 How to release the transport locks Tools 3 mm flat head screwdriver supplied with the instrument Note THERE ARE FOUR TRANSPORT LOCKS SCREWS PRESENT Fig 5 2 A white tag with the red text CAUTION Remove Transport Lock Before Use is fastened to the instrument to attract the user s attention Fig 5 1 The tag in Fig 5 7 and the black arrows in Fig 5 2 indicate where the transport locks are situated Remove the tag and release the transport locks Ensure that the transport locks have been released before you put the instrument into operation Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 15 Chapter 5 Installation Fig 5 1 KingFisher mL transport lock tag present before release of the locks To remove the transport locks unscrew the four screws Fig 5 2 with the 3 mm flat head screwdriver supplied Make sure that the screws are fully unscrewed in the opposite direction 2 Remove the transport lock warning tag Fig 5 7 3 Save the transport lock warning tag for future transportation Ls Thermo Labsystems KingFisher mi Tag attached here Fig 5 2 Location of the transport lo
22. cessor The KingFisher mL instrument Fig 4 5 is designed for maximum 15 tube strips which are compatible with the tip comb The tube strip s is are kept stationary and the only moving assembly is the processing head with tip combs and magnetic rods The head consists of two vertically moving platforms One is needed for the magnetic rods 3 x 5 pieces and the other one for the plastic tip combs Fig 4 4 One tray contains fifteen separate tube strips and one sample processing uses one tube strip containing five tubes One tip comb containing five tips is used for processing five samples at a time Before starting the magnetic particle processing via the keypad and display the samples and reagents are dispensed into the tubes and the tip comb s is are loaded into its their slot s The tube strip s is are placed into the removable tray in the correct position and the tray is pushed into the end position During the operation the front and top lids can be closed Fig 4 7 or open Fig 4 2 Closed lids protect the processing against environmental contamination The operating principle employed is MPP inverse magnetic particle processing technology Fig 4 6 Rather than moving the liquids the magnetic particles are moved from tube to tube containing specific reagents in contrast to the external magnet method Magnetic particles are transferred with the aid of magnetic rods covered with disposable specially designed plastic tip
23. ck screws and warning tag Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 16 Chapter 5 Installation 5 2 2 How to ensure startup 1 Warning Ensure that the mains switch Fig 4 3 on the bottom left of the back panel is in the OFF position 2 Connect the mains supply cable to the ma ns power socket Fig 5 3 at the bottom left of the back panel If you need to use any other type of mains supply cable than supplied use only cables certified by the local authorities Before you plug in the power cable ensure that the voltage on the rating label at the bottom left of the back panel Fig 4 3 corresponds to the local voltage 3 Connect the instrument to a correctly installed line power outlet that has a protective conductor that is grounded connection p Warning Never operate your instrument from a power outlet that has no ground Fig 5 3 Connecting the mains supply cable 5 3 Operational check 1 First switch the instrument ON The instrument performs initialization tests and adjustments The initialization tests are so called self tests 2 The display quickly shows the internal software version and after that the display shows one of the protocols in the internal memory This happens when the initialization tests and adjustments have been completed 3 We recommend that you carry out an empty run for example a demonstration to verify proper instrument operation Thermo Scienti
24. connnnninnnnnnnnnnncnnnnnnnnnnnn nr eren enne 4 3 Principle of magnetic particle processing eesseseseeeee nennen 4 4 Working with a magnetic rod sessssssssssesseeseseeene nennen nnne nnne nnn nnne nnn nns 4 4 1 Collecting magnetic particleS oocoonncccccnanacnccnanannnnnnnnnnncnnnn cnn rcrnnn nn nc enne 4 4 2 Releasing magnetic particles oooocccconccinincccnccnnnnnnnananann cnn nn nn nana rr 4 4 3 Washing magnetic particles sesssssssssssssseeeeeee nennen esent nnn 4 4 4 INCUBATION aure 4 4 5 Changing the volume during the magnetic particle processing sssss 5 Installation iiie roni sau dusute Ge asebuawas aaa a O aa aa ea a Oaa aa Ra Ia ap OAD rg B ga aO 5 1 What to do upon delivery siini iieiea i aa A eiai anaa e a 5 1 1 How to Unpack kiani see et ds eee dt aaa uen 5 1 2 Checking delivery for completeness rrnsrrrrransrrvrnrrnavrrrrrnnvnrsrrnsrnesrennrnesrennvnesvennvnsssennnne 5 1 3 Checking for damage during transport oooocccccnncccncnaninnncnaninnnonanannncnnnnnnnncnnnn a nn rrnnnn narran 5 1 4 Environmental requirements cooccccccccccncnononannncnnnnnnnnnnnnnnn nn ae ener nene 5 1 5 Things TO lt aVOId n la DUE EROR ii a 5 1 6 Technical prerequisites cui c eto eanet 5 2 Setups before you put the instrument into operation eesseseeeeeneenennnnnns 5 2 1 How to release the transport loCkKS oocononccncccnnnncconononccnnnncnnn
25. cy 95 vessel washing buffer Incubating temperature Ambient temperature i e no incubation Keyboard Display Software START STOP two cursor keys LCD Fixed internal protocols loading through the external PC Thermo Scientific KingFisher Software can be used to create and modify protocols Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 25 Chapter 8 Technical Specifications 8 2 Safety specifications The KingFisher mL fulfills the following requirements EN 61010 1 1993 A2 1995 IEC 61010 1 1990 A1 1992 A2 1995 The safety specifications are also met under the following environmental conditions in addition to or in excess of those stated in the operating conditions Altitude up to 2000 m Temperature 5 C 40 C Mains supply fluctuations 10 from nominal Installation category overvoltage category ll according to IEC 60664 1 see Note 1 Pollution degree 2 according to IEC 60664 1 see Note 2 Notes 1 The installation category overvoltage category defines the level of transient overvoltage which the instrument is designed to withstand safely It depends on the nature of the electricity supply and its means of overvoltage protection For example in CAT II which is the category used for instruments in installations supplied from a supply comparable to public mains such as hospital and research laboratories and most industrial laboratories th
26. e expected transient overvoltage is 2500 V for a 230 V supply and 1500 V for a 120 V supply 2 The pollution degree describes the amount of conductive pollution present in the operating environment Pollution degree 2 assumes that normally only nonconductive pollution such as dust occurs with the exception of occasional conductivity caused by condensation Both of these affect the dimensioning of the electrical insulation within the instrument Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 26 Chapter 8 Technical Specifications 8 3 In conformity with the requirements The KingFisher mL bears the following markings Type 701 100 240 Vac 50 60 Hz 80 VA CE mark CSA monogram The KingFisher mL conforms to the following requirements 2006 95 EC Low Voltage Directive 2004 108 EC Electromagnetic Compatibility Directive EMC 2002 96 EC Waste of Electrical and Electronic Equipment Safety performance EN 61010 1 1993 A2 1995 IEC 61010 1 1990 A1 1992 A2 1995 including CENELEC Common Modifications US and CA National differences Installation Category Overvoltage Category Il Pollution Degree 2 EN 61010 1 2001 Ed 2 EMC performance EN 50081 1 1992 Generic emission standard Residential commercial and light industry EN 50082 1 1997 Generic immunity standard Residential commercial and light industry EN 61326 1 1997 A1 1998 Product family standard A2 2001 A3 2003 Te
27. e required service This will ensure a faster return of the instrument to the customer The Thermo Fisher Scientific representative or distributor will take care of sending a complaint form that is the Warranty Claim Technical Sheet to the Thermo Fisher Scientific service department The Warranty Claim Technical Sheet contains a more detailed description of the fault symptom or condition Give all the necessary information to the distributor who will fill in and forward the Warranty Claim Technical Sheet to the Thermo Fisher Scientific service department Check Section 7 4 How to pack for service You will find instructions on how to proceed before shipping the instrument for service Check that any necessary decontamination procedure has been carried out before packing See Sections 9 4 1 and 9 4 1 1 on decontamination Refer also to Section 7 3 Decontamination procedure Ensure that the Certificate of Decontamination as well as the return authorization number RGA are sent with the instrument The Thermo Fisher Scientific service department will keep you up to date with the progress of service and provide you with any further details you might need for example on maintenance serviceability troubleshooting and replacement 9 4 1 Decontamination Decontamination should be performed in accordance with normal laboratory procedures Any decontamination instructions provided with the reagents used should be followed A decontamination pr
28. ea Caution risk of damage to the instrument other equipment or loss of performance or function in a specific application gt gt pi Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 6 Chapter 2 About the User Manual 2 About the User Manual This user manual has been written for the actual end user for example laboratory technician and provides information on the Thermo Scientific KingFisher mL magnetic particle processor including the installation and operating instructions Read the manual in its entirety prior to operating the instrument This user manual has been designed to give you the information you need to e Install the KingFisher mL e Use the KingFisher mL in routine jobs the processing step e Perform basic maintenance procedures This user manual also describes all the features and specifications of the KingFisher mL hardware and on board software Chapter 15 Appendix A Application Protocols for KingFisher mL explains the processing principles and procedures In Chapter 9 Troubleshooting Guide you will find explanations of all error messages and a problem solving guide The user should be familiar with the contents of Chapter 7 on maintenance For warranty and ordering information refer to Chapters 10 Warranty Certificate and 11 Ordering Information In an effort to produce useful and appropriate documentation we appreciate your comments on this document to your local Therm
29. ect the outside of the instrument using a cloth dampened with 70 ethanol b Place the instrument in a large plastic bag Ensure that the lids are open 6 Place a cloth soaked in the prepared solution into the bag Ensure that the cloth does not make contact with the instrument 7 Close the bag firmly and leave the instrument in the bag for at least 24 hours 8 Remove the instrument from the bag 9 Clean the instrument using a mild detergent 10 Remove any stains using 70 ethanol Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 23 Chapter 7 Maintenance 11 After performing this decontamination procedure enclose a signed and dated Certificate of Decontamination both inside the transport package and attached to the outside of the package 7 4 How to pack for service When you ship the instrument for service remember to Inform about the use of hazardous materials Decontaminate the instrument beforehand Install the transport locks Pack the instrument according to the enclosed packing instructions Use the original packaging to ensure that no damage will occur to the instrument during shipping Any damage will incur additional labor charges e Enclose a dated and signed Certificate of Decontamination see Section 9 4 1 1 both inside and attached to the outside of the package in which you return your instrument or other items e Enclose the return authorization number RGA given by the T
30. ed onto the wall of the tips The magnetic rods together with the tip comb s having collected the magnetic particles can be lifted out of the tubes and transferred into the next tubes 4 4 2 Releasing magnetic particles After collection of the magnetic particles the magnetic rods together with the tip comb s are lifted from the tubes the magnetic rods are lifted off and the tip comb s is are lowered into the next tubes containing a reagent Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 12 Chapter 4 Functional Description Magnetic particles are released by moving the tip comb s up and down several times at considerably high speed until all the particles have been mixed with the substance in the next reaction 4 4 3 Washing magnetic particles Washing the magnetic particles is a frequent and an important processing phase Washing is a combination of the release and collection processes in a tube filled with washing solution To maximize washing efficiency the magnetic rods together with the tip comb s are designed to have minimized liquid carrying properties 4 4 4 Incubation To keep the magnetic particle suspension evenly mixed in long running reactions the tip comb s can be moved up and down from time to time 4 4 5 Changing the volume during the magnetic particle processing The volume of the first tube can be larger than the volume of the next tube and this is used for concentration purpos
31. entrifuge tubes should be inserted into the tray so that the caps are so near to the surface of the tray as possible and are all pointing right This allows free tip comb movements 6 4 Shutdown Switch the KingFisher mL off by pressing the power switch Fig 4 3 at the bottom left of the back panel of the instrument into the OFF position Remove any tube strips still on the tube strip tray Dispose of all tube strips as biohazardous waste Remove any tip combs still in the instrument Dispose of tip combs as biohazardous AN waste Wipe the tray surface and the adjacent instrument surface with a soft cloth or tissue paper moistened with distilled water a mild detergent SDS sodium dodecyl sulfate or soap solution If you have spilt infectious agents on the tray disinfect with 70 alcohol or some other disinfectant see Section 7 3 Decontamination procedure Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 21 Chapter 7 Maintenance 7 Waintenance 7 1 Regular and preventive maintenance For reliable daily operation keep the instrument free of dust and liquid spills Abrasive cleaning agents are not recommended because they are likely to damage the paint finish We recommend you clean the case of the instrument periodically to maintain its good appearance A soft cloth dampened in a warm mild detergent solution will be sufficient Clean the outside of the instrument and the tube strip tray with clean
32. er in tube 4 6 The purified sample is released into the elution buffer in tube 5 Volumes 1 Sample particles 1000 ul 2 Washing buffer 1000 ul 3 Washing buffer 1000 ul 4 Washing buffer 1000 ul 5 Elution buffer 200 ul The protocol can be modified according to customer needs Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 41 Appendix A Application Protocols for KingFisher mL A 2 Example of a KingFisher mL total RNA purification protocol The total RNA purification kit which is developed for the purification of total RNA from cultured cells or tissue homogenates can be used with this protocol Normally the kit is based on paramagnetic silica particles that bind RNA in the presence of chaotrophic salts After the binding step the RNA magnetic particle complex is washed extensively in washing buffers to remove any remaining contaminants Finally the purified RNA is released into the elution buffer During the elution step the samples are heated outside the KingFisher mL instrument The approximate processing time for 15 KingFisher mL samples is 40 minutes The KingFisher mL total RNA purification procedure provides high quality RNA for direct use in downstream applications such as RT PCR and Northern blotting You can use ca 2 5 x 10 cells or 30 mg of tissue to purify RNA Yields will vary according to the kit cell and tissue type used in the assay The name of the purification protocol is T
33. eral advantages relating mainly to the principle of operation in that it e Enables automation of complicated manual steps e Enables simultaneous processing and purification e Enables a quicker reaction and a more efficient wash due to the simultaneous shaking during incubation and washing e Facilitates a good collection of bead bound sample due to the efficiency of the magnet e Prevents carry over due to the fact that the same tip is only used for one sample e Facilitates the whole processing with the aid of an internal program Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 8 Chapter 4 Functional Description 4 Functional Description 4 1 Instrument layout 4 1 1 Front view 1 Keypad 2 Display 3 Front lid 4 Top lid Fig 4 1 KingFisher mL front view with lids closed Fig 4 2 KingFisher mL front view with lids open Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 9 Chapter 4 Functional Description 4 1 2 Back view ON OFF switch Mains power supply socket Cooling air outlet Serial port RS 232C hon Fig 4 3 KingFisher mL rear view 4 1 3 Internal view Tube strip tray with tubes Tip comb holder Tip comb slot with tip comb Fixed magnetic rod holder hon Fig 4 4 KingFisher mL internal view Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 10 Chapter 4 Functional Description 4 2 KingFisher mL magnetic particle pro
34. es see Fig 4 7 below i Y T Y 4 y T 4 4 l 4 1 Transter 4 f lj B 1 4 f V 4 il 4 f V 1 4 4 14 4 4 14 4 l A Mi tu A E J v ft 4 4 4 arl J 4 v 4 hu 11 nl te 4 7 f 1 7 y 4 f rv 1 7 Y Jj 4 f f n n Y 4 Y f f q 4 p Y Y AN A J V DI Y 1 4 M My ial f y di y j 4 ry 4 Y f re y y RA S YY 7 VL IL A y Collection of Transfer into Release of Ready magnetic particles a low volume well magnetic particles suspension in a from a large volume low volume well well Fig 4 7 A concentration step during magnetic particle processing Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 13 Chapter 5 Installation 5 Installation 5 1 What to do upon delivery 5 1 1 How to unpack Move the unpacked instrument to its site of operation Unpack the KingFisher mL instrument and accessories carefully with the arrows on the transport package pointing upwards The following notes and instructions are sent with the instrument and are immediately available when you open the package packing instructions packing list Thermo Scientific KingFisher mL Feedback Form Warranty Certificate card KingFisher mL User Manual Caution Do not touch or loosen any screws or parts other than those specially designated in the instructions Doing so might cause misalignment and
35. etic fields direct sunlight draft excessive moisture or large temperature fluctuations e Make sure the working area is flat dry clean and vibration proof and leave additional room for cables lids etc e Make sure the ambient air is clean and free of corrosive vapors smoke and dust e Make sure the ambient temperature range is between 10 C 50 F and 40 C 104 F e Make sure relative humidity is between 10 and 90 non condensing Leave sufficient space at least 10 cm at both sides and at the back of the unit to allow adequate air circulation The KingFisher mL does not produce operating noise at a level that would be harmful No sound level measurements are required after installation Warning Do not operate the instrument in an environment where potentially damaging liquids or gases are present 5 1 5 Things to avoid Do not smoke eat or drink while using the KingFisher mL Wash your hands thoroughly after handling test fluids Observe normal laboratory procedures for handling potentially dangerous samples Use proper protective clothing Use disposable gloves Be sure the working area is well ventilated Never spill fluids in or on the equipment pacemaker or metallic prostheses should not use this product A pacemaker or prostheses may be affected or damaged if it comes in very close contact with a strong magnetic field Warning This product contains strong permanent magnets People wearing a 5 1 6 Techni
36. fic KingFisher mL User Manual Rev 1 3 Cat no 1508260 17 Chapter 6 Routine Operation 6 Routine Operation 6 1 Switching on Before you switch on the KingFisher mL ensure that the voltage on the rating label at the bottom left of the back panel Fig 4 3 corresponds to the local voltage 6 2 How to use the keypad and display The liquid crystal display is a 20 character by 2 line LCD The lower line of the display can show the steps of the protocol while the processing is running The remaining time till the end of the process is shown in the top right corner Furthermore if errors occur they will be displayed on the lower line The keypad and display are shown in Fig 6 7 Thermo Labsystems i KingFisher mL Fig 6 1 KingFisher mL keypad The keys available are two cursor keys or arrow keys direction keys to select the processing step to be run To change to the previous next protocol by scrolling up m To change to the next protocol by scrolling down two function keys START To initiate the processing step STOP To pause terminate the processing step In short STOP paused START the instrument continues after a Pause step STOP paused STOP the processing is terminated Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 18 Chapter 6 Routine Operation 6 3 How to START e First of all slide the tube strip tray out Fig 6 2 Thermo Labsystems KingFisher mL A STAR
37. g materials use the recycling operators known to you For further information contact your local Thermo Fisher Scientific representative Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 24 Chapter 8 Technical Specifications 8 Technical Specifications 8 1 General specifications Technical specifications Overall dimensions ca 290 mm 11 4 in W x 290 mm 11 4 in D x 310 mm 12 2 in H Weight Operating conditions indoor use ca 10 kg 23 Ibs 10 C 40 C RH 90 max Transportation conditions 40 C 70 C packed in transport packaging Storage conditions 25 C 50 C packed in transport packaging Mains power supply 100 240 Vac 50 60 Hz nominal Automatic voltage detection Power consumption 80 VA max 50 VA standby Heat dissipation Computer interface 273 BTU max Serial RS 232C port Baud rate 9600 Character format 1 start bit 8 data bits 1 stop bit no parity Flow control XON XOFF Performance specifications Tube strip tray autoclavable Special design 3 x 5 format Magnetic rods fixed 3 x 5 format Capacity 15 samples run 3 x 5 Vessel volumes 50 ul 1 ml Vessel type disposable Special tube strip 1 x 5 tubes Tip comb disposable Special design 1 x 5 format Magnetic particle size ca gt 1 um Collecting efficien
38. hermo Fisher Scientific representative e Indicate the fault after you have been in touch with your local Thermo Fisher Scientific representative or the Thermo Fisher Scientific service department See Section 8 1 for details on storage and transportation temperatures 7 5 Service contracts We recommend you maintain and service the instrument regularly every twelve months on a contract basis by the manufacturer s trained service engineers This will ensure that the product is properly maintained and gives trouble free service Contact the Thermo Fisher Scientific service department for more details 7 6 Disposal of the instrument Decontaminate the instrument prior to disposal See Sections 7 3 9 4 1 and 9 4 1 1 on decontamination Dispose of the instrument according to the legislation stipulated by the local authorities concerning take back of electronic equipment and waste The proposals for the procedures vary by country Thermo Fisher Scientific has contracted with one or more recycling disposal companies in each EU Member State European Country and this product should be disposed of or recycled through them Further information on Thermo Fisher Scientific s compliance with these Directives the recyclers in your country and information on Thermo Scientific products which may assist the detection of substances subject to the RoHS Directive are available at www thermo com WEEEROHS Regarding the original packaging and packin
39. ication beads cloned DNA cloning DNA DNA complex DNA immobilization DNA purification DNA strand separation DNA template double stranded DNA dsDNA encapsulated magnetic microspheres genome genomic DNA hybride selection KingFisher or Kingfisher magnetic beads magnetic microspheres magnetic particles magnetic separation microspheres nucleotide oligo dT particles PCR PCR amplification PCR product post PCR purification primer probes processing processor purification RNA mRNA RT PCR separation separator silica coated beads silica coated microspheres silica coated particles streptavidin coated beads streptavidin coated microspheres streptavidin coated particles subtraction libraries superparamagnetic beads superparamagnetic microspheres template Thermo Fisher Scientific Thermo Scientific Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 35 Chapter 12 Glossary and Abbreviations 12 2 Literature Aasheim H C Deggerdal A Smeland E B and Hornes E 1994 A simple subtraction method for the isolation of cell specific genes using magnetic mono disperse polymer particles Biotechniques 16 4 716 721 Coche T Dewez M and Beckers M C 1994 Generation of an unlimited supply of subtracted probe using magnetic beads and PCR Nucleic Acids Res 22 1322 1323 Cudjoe K S 1999 Immunomagnetic particle based techniques Overview In Eds Robin
40. ion Protocols for KingFisher ML ooooccnnnnccccnnnocccnncoonononoconnnnnnnannnnnnnnnnoss 41 A 1 Example of a cell or protein purification protocol using KingFisher mL 41 A 2 Example of a KingFisher mL total RNA purification protocol eee 42 16 Appendix B Thermo Scientific KingFisher mL Brief User s Guide 44 17 Appendix C Thermo Scientific KingFisher mL Feedback Form sess 45 18 Appendix D Addresses iere nte rient rra nintendo dox on Marre epu dus 46 19 Appendix E Supplementary Application Protocols rrrrrnnnvnnnnnnnvnnnnnnnvnvnnnnnnvnrennnnvnnnrnnnenn 47 Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 4 Table of Contents Copyright Copyright O 2007 Thermo Fisher Scientific All rights reserved First edition published in 2000 Printed in Finland Reproduction of the accompanying user documentation in whole or in part is prohibited Patents KingFisher mL has national and international patents as well as patents pending Trademarks and Finntip and KingFisher are registered trademarks of Thermo Fisher Scientific Virkon is a registered trademark of Antec International Ltd All other trademarks and registered trademarks are property of their respective holders Disclaimer Thermo Fisher Scientific reserves the right to change its products and services at
41. ired This action does not break the run e Select the name of the method e Use the START key to initiate the run For startup refer to Section 6 1 Switching on Note The samples are collected immediately after the run and transferred into other tubes for longer storage Slip end faces left Fig 6 5 Tube strip tray containing tubes specially designed for the KingFisher mL Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 20 Chapter 6 Routine Operation 6 3 1 Usage of microtube tray in elution The purification protocol can be chosen to use 2 ml microcentrifuge tubes in the elution step Smaller tubes e g 1 7 ml cannot be used as they might get stuck in the tip comb If elution in microcentrifuge tubes is chosen in the KingFisher Software protocol with the KingFisher mL the microtube tray is positioned on the empty KingFisher mL tray tube strip Fig 6 6 Microcentrifuge tubes are inserted and filled with buffer This double tray pack is then inserted into the instrument after removing the previous tray see Fig 6 6 When the instrument pauses and CHANGE TRAY is displayed the first tray is removed from the instrument and the second tray is inserted The protocol continues by pressing the START key The following tube models have been tested and are recommended Elkay 1 9 ml Cat no 000 micr 190 and Costar 2 0 ml Cat no 3213 Fig 6 6 Microtube KingFisher mL double tray Note The microc
42. n devs geet canadian cutie ERE eR FREE E RE EFT ERE caches 18 Hit 7 1 UY q POPE ONE COS CO E MSS 36 M Magnetic particle processor ccccccccecsesesesesecececececececesecesecesauauauauauauauauauaeaeauaeauanauasaaaaaes 7 8 11 30 41 Magnetic particles dices ada 8 11 12 13 30 35 42 collection Oficial di ida idad EE 12 release of O NN 12 Wash fusk A ta RETE eua TET A EE 12 13 Magnetic TOA LR 8 10 11 12 13 22 25 29 30 35 M ns ura arta id 10 23 25 26 Maid TIME 22 24 44 Maintenance cui A ad dav caus DR EVE aan BORN EA YE ia 7 22 24 31 O Operational check dee terc pe cce ete dn eer eer ken Meg Weta o Ye Cr ester tg ds 17 Ordering information s nro rea Poor ai SPA NER ENTERS EET ita iia 7 34 P PACIEN Pde one Merk i AE ob cds 14 24 31 INStEUCtONS a a dada 14 24 ic E E EA EE E R A EE SA I A E E A E Lewes Gamcets 14 materials ii aaa al a ta a aae S a a a aeia 24 PER A A AA N olan A E EAE AESA EA 35 36 Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 38 Chapter 13 Index Pate MEE A AA A EE a AA EAA E it irak 34 CA teie 22 PrOCOSSINO NCAA ida 11 Protocol E EE E AE A A 8 18 30 31 36 41 42 44 R Releasing the transport lOCKS aiani cree diede e E aa ANE ai 15 RNA var Fun nia TEATE AA ita EE A E TEET 30 35 36 42 43 ugue duum 35 S Samples ore oer rore ees 8 11 15 19 20 23 25 30 34 35 41 42 43 44 SUI IEEE 24 25 29 31 34 CONTaCtSuaakdanu EPISOD
43. nnnnnnnn nc nn entree nennen enne 5 2 2 How to ensure StaltUP cccccccccnnnnnononnnnnnnnnnnnnnnnnnnn rn nn nana rre nenes 53 Operational check dieere A A A enti AS 6 Routine Operation inii irri cid ee i 61 Switching OM zii eer nn eni Y reker vedsiden 6 2 How to use the keypad and display sssssssssssssesee eene enne nnne 6 3 Howto STAR Tiida ii 6 3 1 Usage of microtube tray in elution oooonncccnnonicccnnononcnnnnononn nono nnnn cnn rra rra rra 64 SHUTO ariannin ADA Be MIAINtOM LIII 7 1 Regular and preventive maintenance ococccccccccnnnnnnnnncnnnnnnnnnnnnnn nc nn rn nn nn enn rre 7 1 1 How to clean the tube strip tray occccccononoconcnnnoncnancnnnnnnnnncnnnnnnnnnnn nc nn nn eee eee nn nnr nn 7 1 2 How to clean the magnetic TOS ccoccccccnnonononcninnnnnnnnnnnnn cnn nen n nan naar ae nn rre 7 1 3 How to clean the processing chamber sss 7 2 Disposal of materials intei einen e en n em equ n R d eR RES RE AD RE a a e RU dne 7 3 Decontamination procedure cccccccnnnnnnnnnnnncnnnnnnnnnnnn nn n nr nn rre 74 Howto Pack fOr SEIVICO cialis e Ene rie E ERE eua 7 5 Service contracts iaa 7 6 Disposal of the instrument eene nennen enne ener nnne nn nnns Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 Table of Contents 8 Technical Specifications iii nn 25 81 General specifiCatiOlis ceceeie cual fin ce 0E NN oe a e A EE EA A A E 25 8
44. o Fisher Scientific representative Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 7 Chapter 3 Introduction to the KingFisher mL 3 Introduction to the KingFisher mL 3 1 Intended use The KingFisher mL magnetic particle processor Fig 3 1 is intended for professional research use by trained personnel The instrument is intended for automated transfer and processing of magnetic particles in a 1 ml tube scale Use for self testing is excluded It is recommended that Good laboratory Practices GLP are followed to guarantee reliable analyses 3 2 Principle of operation The KingFisher mL magnetic particle processor Fig 3 7 is designed for the automated transfer and processing of magnetic particles in a 1 ml tube well scale The principle of the KingFisher mL system patents pending is based on the use of magnetic rods covered with disposable specially designed tip combs and tube strips The instrument functions without any dispensing or aspiration parts or devices Samples and reagents including magnetic particles are dispensed into the tubes according to the corresponding kit instructions The steps of the protocol that is selected by the user via the keypad and display have already been preloaded in the on board software Fig 3 1 KingFisher mL magnetic particle processor 3 3 Advantages of processing magnetic particles with the KingFisher mL The KingFisher mL magnetic particle processor provides sev
45. ocedure is only recommendable when infectious substances have been in direct contact with any part s of the instrument Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 31 Chapter 9 Troubleshooting Guide 9 4 1 1 Certificate of Decontamination The decontamination procedure is required prior to shipping the instrument to Thermo Fisher Scientific Oy e g for repair If for any reason the instrument is shipped back to Thermo Fisher Scientific Oy it must be accompanied by a dated and signed Certificate of Decontamination which must be attached to the outside of the package containing the instrument See Section 7 3 Decontamination procedure Failure to confirm decontamination will incur additional labor charges or at worst the items will be returned for proper cleaning Before returning any instrument s or item s ensure that they are fully decontaminated Confirm A or B status Name Address Tel Fax Name Serial no A I confirm that the returned items have not been contaminated by body fluids toxic carcinogenic or radioactive materials or any other hazardous materials B I confirm that the returned items have been decontaminated and can be handled without exposing the personnel to health hazards Materials used in the unit Chemicals Biological e Radioactive Specific information about contaminants Decontamination procedure Date and place
46. otal_RNA_mL_1 Pipetting the reagents KingFisher mL Tube no KingFisher mL tube strip 1000 ul cell lysate OR tissue homogenate 50 ul magnetic particles Washing buffer 1 1000 ul Washing buffer 2 1000 ul Washing buffer 2 1000 ul Elution buffer 200 ul Protocol Total RNA mL 1 The sample is incubated with magnetic particles in tube 1 for 19 minutes The particles are washed in washing buffer 1 in tube 2 The particles are washed in washing buffer 2 in tube 3 The particles are washed in washing buffer 2 in tube 4 The particles are released into the elution buffer in tube 5 The protocol is paused for the heating step The text marked PAUSED appears on the screen The tray is removed OR the samples are transferred into microtubes for the heat incubation 8 After incubation the tray is returned to the KingFisher mL OR the samples are transferred back into tube 5 The protocol will continue after you press START 10 The particles are removed and returned into tube 4 DIRON N Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 42 Appendix A Application Protocols for KingFisher mL Sample preparation from cultured cells Transfer the cultured cells suspended in PBS into a suitable centrifuge tube Collect the cells by centrifugation for 5 minutes at 6000 rpm and discard the supernatant If frozen cells are used thaw the cells on ice centrifuge the cells for 5 minutes a
47. p the RNA yield drastically Note f a rotor stator homogenizer is not available other homogenization methods can be used instead For example soft tissues such as liver can be homogenized using for example a pestle and needle syringe method Always follow the centrifugation instructions above despite the homogenization method used Add up to 1000 ul of the supernatant into the KingFisher mL tube according to the instructions Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 43 Appendix B Thermo Scientific KingFisher mL Brief User s Guide 16 Appendix B Thermo Scientific KingFisher mL Brief User s Guide e Switch the KingFisher mL on p 17 e Pipette the samples reagents and wash buffers into the tube strips p 19 41 e Load the tray s onto the instrument p 19 e Insert the tip comb s into the tip comb holder slot s p 19 e Select the desired protocol from the keypad and display p 18 e Press the START button p 19 e After the end of the run remove the tube strips and tip combs p 21 e Maintain your KingFisher mL instrument regularly p 22 Thermo Labsystems e KingFisher mL Fig 16 1 KingFisher mL keypad Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 44 Appendix C Thermo Scientific KingFisher mL Feedback Form Please send to Thermo Fisher Scientific Oy Fax 358 9 2910415 17 Appendix C Thermo Scientific KingFisher mL Feedback Form
48. parameters lost The track cannot be moved because the head is not up The calculated head position does not agree with the head home sensor signal The head has not arrived at the home sensor within the allowed time The calculated track position does not agree with the track home sensor signal The track has not arrived at the home sensor within the allowed time The calculated magnetic rod position does not agree with the rod home sensor signal The magnetic rods have not arrived at the home sensor within the allowed time The nonvolatile parameters of the instrument are lost Press STOP If the fault cannot be remedied then switch the power on and off Check that there is nothing preventing the head from moving up and down Check that there is nothing preventing the head from moving up and down Check that there is nothing preventing the track from moving left or right Check that there is nothing preventing the track from moving left or right Check that there is nothing preventing the magnetic rods from moving up and down Ensure that the tip comb is fully in Check that there is nothing preventing the magnetic rods from moving up and down Contact authorized service Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 29 Chapter 9 Troubleshooting Guide 9 3 Frequently asked questions FAQ about KingFisher mL processing Q1 What does the KingFisher
49. rrrnnvrrnnnnnvnnnnnnnvnnnnnnnnvnrnnnnnvnnnrnnnenn 41 16 Appendix B Thermo Scientific KingFisher mL Brief User s Guide 44 17 Appendix C Thermo Scientific KingFisher mL Feedback Form sess 45 18 Appendix D Addresses ernnnnnnnnnnnnvnvnnnnnnnnnnnnnvnnnnnnnnnnnnannnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnennnnnnnnnnnnnnn 46 19 Appendix E Supplementary Application Protocols rrrnnnnnvnnnnnnnvnnnnnnnvnvnnnnnnvnnnrnnnvenernnneen 47 Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 2 Table of Contents Table of Contents 1 Safety Symbols and Markings xvrrrrnnnnvnnnnnnnvnnnnnnnnvnnennnnvnnennnnvnnennnnvnnennnnvnnennnnennnensnnnnrennnnennennnne 2 About the User Manual dean Aa Kendra e Sepa coins 3 Introduction to the KingFisher mL arnnnennnnnnnnnnnnvnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnen 3 1 Intended Use cid ai 3 2 Principle ot operational e enit Add dee A Aa da ade 3 3 Advantages of processing magnetic particles with the KingFisher mL 4 Functional Description deiecit naeh ilatina vendes dani 4 1 Irnistr ment AV OUTS coc eoo eco Al to eee e a redacto nudius 4 1 1 Front VIeW uus invo n E oer EUM TR SEIN 4 1 2 Back view ze set ce ese ae td 4 1 3 Internal VIew idu e RE HO T ERR a 4 2 KingFisher mL magnetic particle proCesSOF ccccnnono
50. son R K Batt C A and Patel P D Encyclopaedia of Food Microbiology Academic Press London UK Pp 1088 1094 Hornes E and Korsnes L 1990 Magnetic DNA hybridisation properties of oligonucleotide probes attached to superparamagnetic beads and their use in the isolation of poly A mRNA from eukaryotic cells Genet Anal Tech Appl GATA 7 145 150 Jakobsen K S Breivold E and Hornes E 1990 Purification of mRNA directly from crude plant tissues in 15 minutes using oligo dT microspheres Nucleic Acids Res 18 3669 Jakobsen K S Haugen M S boe Larsen S Hollung K Espelund M and Hornes E 1994 Direct mRNA isolation using magnetic Oligo dT Beads A protocol for all types of cell cultures animal and plant tissues In Eds Uhl n M Hornes E and Olsvik Advances in Biomagnetic Separation Eaton Publishing Pp 61 71 Lambert K N and Williamson V M 1993 DNA library construction from small amounts of RNA using paramagnetic beads and PCR Nucleic Acids Res 21 775 776 Lee Y H and Vacquier V D 1992 Reusable cDNA libraries coupled to magnetic beads Anal Biochem 206 206 207 Rudi K Kroken M Dahlberg O J Deggerdal A Jakobsen K S and Larsen F 1997 Rapid universal method to isolate PCR ready DNA using magnetic beads BioTechniques 22 506 511 Sinclair B 1998 To bead or not to bead Applications of magnetic bead technology Scientist 12 16 19
51. st standards Performance limits EN 55022 1998 Class B 150 kHz 1 GHz EN 61000 3 2 1995 A1 1998 Class A A2 1998 A13 1997 A14 2000 EN 61000 3 3 1995 EN 61000 4 2 1995 A1 1998 4 kV CD 8 kV AD Criteria B EN 61000 4 3 1996 A1 1998 3 V m 80 MHz 2 GHz Criteria A EN 61000 4 4 1995 1 kV Criteria B EN 61000 4 5 1995 2 kV line to ground 1 kV line to line Criteria B EN 61000 4 6 1996 3 Vims 150 kHz 80 MHz Criteria A EN 61000 4 11 1994 30 10 ms Criteria B 60 100 ms Criteria C 100 20 ms Criteria B gt 95 5 s Criteria C Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 27 9 Troubleshooting Guide 9 1 Error messages and warnings Error message Warning Command error Cannot move track Head position error Head timeout Track position error Track timeout Magnets pos error Magnets timeout Serial number error Serialno already set Too high pos offset Invalid parameter Out of memory No acceleration tbl NV parameters lost Progmem full Time already expired Chapter 9 Troubleshooting Guide Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 28 9 2 Troubleshooting guide Error Cause Chapter 9 Troubleshooting Guide Action e Cannot move track e Head position error e Head timeout e Track position error e Track timeout e Magnets position error e Magnets timeout e Nonvolatile
52. t 1000 rpm and discard the supernatant Add lysis buffer including 10 ul B ME 1000 ul buffer so that the final concentration ranges from 2 5 x 10 cells 1000 ul Mix well by vigorously pipetting up and down several times until the lysate is homogenous Vortex for an additional 30 seconds at full speed Note Ifa lot of foam has formed keep the sample on ice while filling the KingFisher mL tube strips with other reagents or centrifuge the sample for 1 minute at 1000 g 3000 rpm in a microcentrifuge Note If a centrifuge is used ensure that you follow the centrifugation instructions Do not exceed the time and speed as this will drop the RNA yield drastically Mix the homogenate gently by pipetting and add the cell mixture into the KingFisher mL tube strip according to the instructions Sample preparation from tissue Weigh the needed amount of tissue up to 30 mg and add 900 ul of lysis buffer including 10 ul B ME 1000 ml buffer B ME Depending on the homogenizer and the tool used change the amount of tissue and lysis buffer using the same ratio as above see the instructions in the previous section Homogenize the sample on ice in the presence of lysis buffer for 5 90 seconds depending on the toughness of the tissue Centrifuge the homogenate for 1 minute at 1000 g 3000 rpm in a microcentrifuge Note Ensure that you follow the centrifugation instructions Do not exceed the time and speed as this will dro
53. will void the instrument warranty To lift the instrument put your fingers under the bottom on either sides and lift it with your back straight Retain the original packaging for future transportation The packaging is designed to assure safe transport and minimize transit damage Use of alternative packaging materials may invalidate the warranty Also retain all instrument related documentation provided by the manufacturer for future use 5 1 2 Checking delivery for completeness Check the enclosed packing list against order If any parts are missing contact your local Thermo Fisher Scientific representative or Thermo Fisher Scientific Oy 5 1 3 Checking for damage during transport Visually inspect the transport package the instrument and the accessories for any possible transport damage If the carton has been damaged in transit it is particularly important that you retain it for inspection by the carrier in case there has also been damage to the instrument Visually check all interconnections in the basic instrument Check that there are no loose parts inside the instrument If any parts are damaged contact your local Thermo Fisher Scientific representative or Thermo Fisher Scientific Oy Thermo Scientific KingFisher mL User Manual Rev 1 3 Cat no 1508260 14 Chapter 5 Installation 5 1 4 Environmental requirements When you set up your KingFisher mL avoid sites of operation with excess dust vibrations strong magn

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