Product Manual S 400 13 L
Contents
1. 8 3 3 Self programming of the KingFisher Flex instrument eere 10 4 Typical Results 2 iionieieeiinn ia C nan neon i ia n rr ei ee cea Dira id 12 4 1 PUN ILY 12 5 Troubleshooting cceseeeeeeeeeeeeeneeeeeeeseeneeuseneeeeaseeeeeesseeeeaseseeeeaseeeeeeaseneeeaseeeeeasseeneenseneeenees 13 DNA NDNIIDDqm 13 7 Warranty and Disclaimer of Liability eese 14 8 Supplementary Notes cnni tenerte aient Darii eden ees eei iav uaar 14 8 1 Ordering Informatior 1 ansasen ansaan Derretiedeeeet ac eseor i esc cn ice eae c esie 14 OEICO Tur Co 14 8 3 Contact and Suppott 5 eire ettari eie EReOE eios dria tdi a ee aa e dee tid cuam eque Us 14 9 Change INDO X qe iSi 15 1 Kit Components BIOTECON Diagnostics Kit Contents Bottle Tube Label Contents Function consumable Bottle No 1 foodproof Magnetic Preparation 3x 250 ml Kit III Lysis Buffer For lysis of cells and extraction of DNA Bottle No 2 foodproof Magnetic Preparation e 2 x 24 ml add 2 x 56 ml absolute isopropanol Kit III Binding Buffer For binding of DNA to the magnetic beads Bottle No 3 foodproof Magnetic Preparation2. 3 Storage of Samples If you want to Then Continue Use the eluted DNA directly The eluted DNA can be stored for several weeks at 4 8 C or stored at Stop 20 C for long term storage 3 3 Self programming of the KingFisher Flex instrument Protocol information Protocol name foodproof MPK Ill Kit name foodproof MPK III Description KingFisher Flex protocol for isolation of genomic DNA from raw material and food products of plant and animal origin Plate layouts Binding Plate Microtiter 96 Deep Well Plate Name Well volume pl Total reagent volume pl Type Sample 500 Sample Binding Buffer 250 Reagent Magnetic Beads 20 Reagent Washing Plate Microtiter 96 Deep Well Plate Name Well volume pl Total reagent volume pl Type Wash Buffer 600 Reagent Washing Plate Il Microtiter 96 Deep Well Plate Name Well volume pl Total reagent volume pl Type Wash Buffer II 800 Reagent Washing Plate Ill Microtiter 96 Deep Well Plate Name Well volume ul Total reagent volume pl Type Wash Buffer II 800 Reagent Elution Plate Microtiter 96 Well Plate Name Well volume pl Total reagent volume pl Type Elution Buffer 150 Reagent Tip Plate Microtiter 96 Well Plate Name Well volume pl Total reagent volume pl Type 10 fa c A c G Cc A T c c c c T T c A T A T c T T c c A T A c c A File Steps
3. DM det P3 I S BIOTECON Diagnostics For food testing purposes FOR IN VITRO USE ONLY foodproof Magnetic Preparation Kit Il Version 3 July 2015 For the automated isolation of DNA from raw material and food products of plant and animal origin using the KingFisher Flex instrument Order No S 400 13 L Kit for 5 x 96 isolations Store the Magnetic Beads at 2 8 C Store lyophilized Proteinase K at 2 8 C Store all other kit components at room temperature A T ES BIOTECON Diagnostics Table of Contents 1 enu udi cree 3 Kit COMtemts secs cscdeoee ieee aa ee ee aaa 3 Chemical HazZard ccccccccssscceceenceeeseeeeeeseaeeeeseneeeescaeeesscaaececeneeeesscaeeesseeeeeeseneeeeseneeeeseaes 4 Number of Preparations ccccccc sccccteccecceg se leneveecenedbgecgeneseeecuestececgnucrenessccpeessseeneedueddesnssesinenstiee 5 LOA eva EET 5 2 How to Use this Product cccscceeessseeneseeeeeseseeeeeasseeeeaseeeeeeaseeeseasseneeasseeeeeaseeeeeesseeeensenne 5 ANLE Ja c 5 deemed 5 ELI 6 Sample A CUCIEIN eds 6 Quality COM 6 3 Procedures and Required Materials esee esee eene 6 3 1 Before you beglh cicero nana nnb e Dn ie ER a da li eed da E E EENE 6 Preparation of Kit Working Solutions nennen 6 3 2 irte m
4. and Wash Buffer II bottle to indicate the addition of ethanol Low amount Beads of Magnetic Mix the Magnetic Beads thoroughly before pipetting to the Binding Plate DNA does not perform well in real time PCR Salt carryover during elution Check the Wash Buffers for salt precipitates If there are any precipitates solve these precipitates by careful warming Ensure that Wash Buffers are stored at room temperature Low A260 A280 ratio from UV measurement eluted DNA is brown colored Small part of the magnetic particles are left in the elution Centrifuge at full speed for 1 min and transfer supernatant contains DNA to a new tube 6 Appendix KingFisher Flex Software The KingFisher Flex Software was used to create assay files for the KingFisher Flex instrument The respective assay file can either be transferred onto the KingFisher Flex workstation or be started directly from within the Bindlt software Keep in mind that assay files run directly will not be stored in the workstation memory 13 ES BIOTECON Diagnostics 7 Warranty and Disclaimer of Liability Limited Warranty and Disclaimer of Liability BIOTECON Diagnostics warrants that this product is free from defects in materials and workmanship through the expiration date printed on the label and only if the following conditions are complied with 1 The product is used according to the guidelines and instructions set forth in
5. the product literature 2 BIOTECON Diagnostics does not warrant its product against any defects when defect is a result of material or workmanship not provided by BIOTECON Diagnostics defect is caused by misuse or use contrary to the instructions supplied the product is contaminated by improper storage or handling 3 All warranties of merchantability and suitability for a particular purpose written oral expressed or implied shall extend only for a period of one year from the date of manufacture There are no other warranties that extend beyond the conditions described here 4 BIOTECON Diagnostics does not assume responsibility to any purchaser of its product for any undertaking representation or warranty made by any dealers or distributors selling its products beyond those herein expressed unless deviating terms are expressed in writing by an officer of BIOTECON Diagnostics 5 BIOTECON Diagnostics does not assume responsibility for incidental or consequential damages including but not limited to responsibility for loss of use of this product removal or replacement labor loss of time inconvenience expense for telephone calls shipping expenses loss or damage to property or loss of revenue personal injuries or wrongful death 6 BIOTECON Diagnostics reserves the right to replace or allow credit for any modules returned under this warranty 8 Supplementary Notes 8 1 Ordering Information Order No S 400 13 L Please send yo
6. C A T C C G T A T Cc T G 0001111 BIOTECON Diagnostics A TC CAT CC CT AT CT G SS ey Eluting Elution Plate Beginning of step Precollect No Release time speed 00 00 10 Medium Mixing heating Mixing time speed 00 10 00 Medium Heating temperature C 90 Preheat No End of step Postmix No Collect count 4 Collect time s 3 ReleaseBeads1 Washing Plate Ill 959 Release time speed 00 00 10 Fast Sp Leave Tip Plate 4 Typical Results 4 1 Purity Purified DNA is free of other cellular components and DNA polymerase inhibitors 12 5 Troubleshooting ES BIOTECON Diagnostics Problem Possible Cause Recommendation Low DNA yield or Kit stored under non optimal e Store the Magnetic Beads at 2 8 C purity conditions Store lyophilized Proteinase K at 2 8 C Store diluted Proteinase K at 20 C Store all other kit components at room temperature Buffer or other reagents were exposed to conditions that reduced their effectiveness Store all buffers at room temperature Close all reagent bottles tightly after each use to preserve pH and stability and to prevent contamination After lyophilized reagents are reconstituted store at 20 C Ethanol not added to Wash Buffer or Wash Buffer II Add absolute ethanol to the Wash Buffer and Wash Buffer Il before using After adding ethanol mix the Wash Buffer and Wash Buffer II well and store at room temperature Always mark the Wash Buffer
7. e 3 x 80 ml add 3 x 80 ml absolute ethanol Kit III Wash Buffer For removing impurities Bottle No 4 foodproof Magnetic Preparation e 5 x 60 ml add 5 x 140 ml absolute ethanol Kit III Wash Buffer II For removing impurities Bottle No 5 foodproof Magnetic Preparation e 110 ml Kit III Elution Buffer For elution of DNA Tube No 6 foodproof Magnetic Preparation e 20 x lyophilized Kit I Proteinase K working e For protein digestion and inactivation of endogenous solution nucleases Dilute Proteinase K by addition of 1 8 ml of ddH2O mix thoroughly and store like described below Tube No 7 foodproof Magnetic Preparation e 10 x 1 1 ml Kit III Magnetic Beads For binding of DNA Consumable Deep Well Plate 5 x 4 plates Consumable Elution Plate 5 x 2 plates Elution Plates and Tip Plates are identically Use one provided Elution Plate as a Tip Plate Consumable Tip Comb 96 DWH 5x1 comb Consumable Adhesive Seal 5 x 2 foils d Hn EM E S BIOTECON Diagnostics Chemical Hazard When working with chemicals always wear a lab smock disposable gloves and protective goggles Observe the legal requirements for working with biological material For more information please consult the corresponding material safety data sheet MSDS If any reagent bottles are damaged or leaky wear gloves and protective goggles when discarding the bottles to avoid injuries European Community risk and safety phrases for
8. e material Sample cross contamination and reagent cross over is effectively eliminated by the provided purification assay The KingFisher Flex instrument uses magnetic rods to transport the DNA bound to magnetic particles through the various purification phases binding washing and elution The volume of buffers and other liquids necessary for DNA isolation is reduced to a minimum Eliminating the direct liquid handling and increasing the automation level results in a fast reliable and robust technique To achieve an efficient lysis and high DNA yields the samples are first lysed with optimized lysis buffer and Proteinase K in a separate step After lysis the DNA binds to the magnetic beads whereas contaminations metabolites and enzyme inhibitors are efficiently removed during the following three wash steps Finally highly purified DNA is eluted in Elution Buffer The purified high quality DNA is ready to use for following downstream applications like PCR or can be stored at 20 C for subsequent use BIOTECON Diagnostics Basic Steps Step Description Sample lysis by incubation with the foodproof Magnetic Preparation Kit III Lysis Buffer and foodproof Magnetic Preparation Kit III Proteinase K 2 DNA is bound to magnetic beads 3 Washing of bound DNA to remove proteins and other cellular impurities 4 Purified DNA is recovered using the Elution Buffer Sample Material 200 mg sample material from raw
9. ed S 400 13L 20 4 BIOTECON Diagnostics GmbH Hermannswerder 17 14473 Potsdam Germany Phone 49 0 331 2300 200 Fax 49 0 331 2300 299 www bc diagnostics com bcd bc diagnostics com 15
10. ll provide the next free loading position automatically When all plates are loaded press START again to initialize the program ES BIOTECON Diagnostics The following purification steps will run automatically on the KingFisher Flex System Binding of the DNA Automatically sample mixing for 5 min Magnetic Beads separation Transfer of the magnetic particles to Washing Plate First Washing Automatically sample mixing for 1 5 min Magnetic Beads separation Transfer of the magnetic particles to Washing Plate II Second Washing Automatically sample mixing for 1 min Magnetic Beads separation Transfer of the magnetic particles to Washing Plate III Third Washing Automatically sample mixing for 1 min Magnetic Beads separation Drying Drying of the Magnetic Beads outside Washing Plate Ill for 5 minutes Transfer of the Magnetic Beads to the Elution Plate Elution of the DNA Incubation of magnetic particles in the Elution Buffer for 10 minutes at 90 C by continuously mixing Magnetic Beads separation The Magnetic Beads will automatically be removed and transferred in Washing Plate III disposal Note 1 After finishing the extraction protocol the Elution Plate contains the extracted DNA 2 If the extracted DNA contains carryover of magnetic particles transfer the DNA into a 1 5 ml reaction tube and centrifuge at maximum speed for 1 minute Transfer the clear supernatant contains the DNA into a new tube
11. ls decontaminate work surfaces and use a biosafety cabinet whenever aerosols might be generated The following protocol describes the automated DNA isolation from 200 mg sample material with the KingFisher Flex instrument Note Transfer up to 200 mg homogenized sample material into a 2 ml reaction tube and add 1400 ul of Lysis Buffer and 70 ul of Proteinase K Incubate for 30 min at 65 C with shaking in a thermomixer Centrifugate for 10 min at 12 000 g Switch on the KingFisher Flex instrument Before starting the purification process with the KingFisher Flex instrument please read carefully the user manual Resuspend Vortex the Magnetic Beads thoroughly directly before use Tip Plate Place the Tip Comb 96 DWH on a Tip Plate Use one provided Elution Plate as Tip Plate These are identical Prefill the Binding Plate the Washing Plates and the Elution Plate as described below Binding Plate Add 250 yl Binding Buffer and 20 ul Magnetic Beads Washing Plate I Add 600 ul Wash Buffer I Washing Plate Il Add 800 ul Wash Buffer II Washing Plate Ill Add 800 ul Wash Buffer Il Elution Plate Add 200 yl Elution Buffer Transfer 500 ul of the lysate into the Binding Plate Choose assay file foodproof MPK III on instrument and press START Follow instructions on the instruments display and load the prefilled buffer plates in the right position Confirm with START after each loading step the instrument then wi
12. material and food products from plant and animal origin are processed with the kit Quality Control Each component of the Magnetic Preparation Kit Ill has been tested against predetermined specifications to ensure consistent product quality 3 Procedures and Required Materials 3 1 Before you begin Preparation of Kit Working Solutions Before starting a run bring all reagents to room temperature Gently mix and redissolve any precipitates by warming up to 30 C Swirl gently to avoid foaming foodproof Magnetic Preparation Kit III Lysis Buffer Binding Buffer and Elution Buffer are ready to use Add the required amount of ddH5O to the reaction tube containing the foodproof Magnetic Preparation Kit III Proteinase K Vortex for 5 s and keep the solution on ice Store unused and diluted foodproof Magnetic Preparation Kit IIl Proteinase K at 20 C eo BIOTECON Diagnostics Bottle Preparation of working solution Storage and stability Add 56 ml absolute isopropanol to the bottle Binding Buffer St f ore at room foodproof Magnetic Mix thoroughly and always keep the temperature Stable Preparation Kit Ill Binding bottle firmly closed until the expiry date Buffer Note Check the box on the label of printed on kit label the bottle after isopropanol has been added Add the date for verifiability Add 80 ml absolute ethanol to the bottle Wash Buffer I foodproof Magnetic Mix thoroughly and always keep the Preparati
13. mr Tip 1 md A T c c A A T c G C G T A T T c C G c c T A T c c T A T Pick Up Binding Beginning of step Mixing heating End of step Washing 1 Beginning of step Mixing heating End of step Washing 2 Beginning of step Mixing heating End of step Washing 3 Beginning of step Mixing heating End of step Dry1 A n o A T n Tip Comb 96 DWH BIOTECON Diagnostics Tip Plate no o Binding Plate Precollect Release time speed Mixing time speed Heating during mixing Postmix Collect count Collect time s Washing Plate Precollect Release time speed Mixing time speed Heating during mixing Postmix Collect count Collect time s Washing Plate II Precollect Release time speed Mixing time speed Heating during mixing Postmix Collect count Collect time s o Washing Plate Ill Precollect Release time speed Mixing time speed Heating during mixing Postmix Collect count Collect time s Washing Plate Ill Dry time Tip position 11 n 4 No 00 00 10 Fast 00 05 00 Medium No No 5 10 No 00 00 10 Fast 00 01 30 Medium No No 4 3 No 00 00 10 Fast 00 01 00 Medium No No 4 3 No 00 00 10 Fast 00 01 00 Medium No No 4 3 00 05 00 Outside well tube 4 A C A T C G T A T C C A T c c c T A T C C A T C A T C C C T A T C G C T T C C A T Y Ge C T T
14. on Kit III Wash bottle firmly closed Store at room temperature Stable until the expiry date Buffer Note Check the box on the label of printed on kit label the bottle after ethanol has been added Add the date for verifiability Add 140 ml absolute ethanol to the bottle Wash Buffer II Store at room foodproof Magnetic Mix thoroughly and always keep the temperature Stable Preparation Kit III Wash bottle firmly closed until the expiry date Buffer II Note Check the box on the label of printed on kit label the bottle after ethanol has been added Add the date for verifiability foodproof Magnetic Dissolve Proteinase K by addition of Preparation Kit III 1 8 ml double distilled water ddH2O Proteinase K per reaction tube needed Store at 20 C Stable for 12 months Additional Equipment and Reagents required KingFisher Flex instrument Measuring cylinder 250 ml Pipette and pipette tips Disposable gloves ddH O Vortexer absolute Ethanol 96 98 95 absolute Isopropanol 96 98 95 2 ml reaction tubes Thermomixer for 2 ml reaction tubes e g Eppendorf ThermoMixer comfort ES BIOTECON Diagnostics 3 2 Protocol Caution Always wear gloves during the procedure and follow safety precautions to minimize contact when handling Follow generally applicable safety precautions regulating the work with biohazard materials Properly dispose of all contaminated materia
15. the components of the foodproof Magnetic Preparation Kit IIl to which they apply are listed below Lysis Buffer Proteinase K danger danger H302 319 P305 351 338 H315 319 334 335 P280 305 351 338 310 405 Binding Buffer Wash Buffer danger warning H225 319 336 P210 233 305 351 338 H302 312 332 412 EUH032 P273 H315 Causes skin irritation H319 Causes serious eye irritation H334 May cause allergy or asthma symptoms or breathing difficulties if inhaled H335 May cause respiratory irritation H225 Highly flammable liquid and vapor H336 May cause drowsiness or dizziness H302 Harmful if swallowed H312 Harmful in contact with skin H332 Harmful if inhaled H412 Harmful to aquatic life with long lasting effects EUH032 Contact with acids liberates very toxic gas P280 Wear protective gloves protective clothing eye protection face protection P305 P351 P338 If in eyes Rinse cautiously with water for several minutes Remove contact lenses if present and easy to do Continue rinsing P310 Immediately call a POISON CENTER or doctor physician P405 Store locked up P210 Keep away from heat sparks open flames hot surfaces No smoking P233 Keep container tightly closed P273 Avoid release to the environment 4 ES BIOTECON Diagnostics Number of Preparations 5 x 96 isolations Storage All buffers and kit components of the foodproof Magnetic Preparation Kit Ill except Proteinase K and Magne
16. tic Beads should be stored at room temperature and are stable for at least 12 months under these conditions Proteinase K Lyophilized Proteinase K should be stored at 2 8 C Dissolved Proteinase K must be stored at 20 C Avoid multiple freezing and thawing cycles of diluted Proteinase K Magnetic Beads The magnetic particles should be stored at 2 8 C Wash Buffer and II Wash Buffers charged with ethanol should be stored at room temperature and must be sealed accordingly If any precipitates are visible within the provided solutions solve them by carefully heating up to 30 C room temperature RT is defined as range from 15 30 C 2 How to Use this Product 2 1 Product Overview Test Principle The foodproof Magnetic Preparation Kit Ill in combination with the KingFisher Flex instrument provides fully automated purification of total genomic DNA from up to 200 mg from raw material and food products of plant and animal origin The DNA isolation process is based on patented magnetic bead technology which relies on the interaction of nucleic acids with coated magnetic particles under suitable buffer conditions The kit provides high quality DNA which is suitable for direct use in PCR applications The KingFisher Flex instrument performs all steps of the DNA purification procedure automatically The procedure requires only minimal interaction by the user namely the initial loading of the system and the preparation of the sampl
17. ur order directly to order bc diagnostics com or call us at 49 0 331 2300 200 BIOTECON Diagnostics offers a broad range of reagents and services For a complete overview and for more information please visit our website at www bc diagnostics com 8 2 Trademarks foodproof is a trademark of BIOTECON Diagnostics GmbH KingFisher is a registered trademark of Thermo Fisher Scientific Other brand or product names are trademarks of their respective holders 8 3 Contact and Support If you have questions about this or any other product of BIOTECON Diagnostics please contact our Technical Support contact details at www bc diagnostics com Our expert scientists will provide any request with rapid and effective help We also welcome you to contact us if you have suggestions for enhancing our product performance or using our products in new or specialized ways 14 c c A UT OC 9 Change Index Version 1 August 2013 First version of the package insert Version 2 April 2014 A 2s 7 HB aH C C C A C G C T eu c io BIOTECON Diagnostics Page 4 Change in volume of Proteinase K and double distilled water ddH2O added Page 8 Change in volume double distilled water ddH2O added Page 9 Change in volume of Proteinase K added Version 3 July 2015 Page 3 Change in volume double distilled water ddH2O added Page 7 Change in volume double distilled water ddH2O added Page 8 Change in volume of Proteinase K add
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