- Diagenode
Contents
1. Shutting down the IP Star9 iiiiiiisisssslle lehren 21 icc r ee ee eee 22 Je eitisiizcanpimECHI Pmmm 23 lechnical ASSISIONCO P PIP m 23 Ordering MTOrmatOn sao50 che eee beens eek eee reek EA wee oe Back Cover Diagenode Inc North America Phone 1 862 209 4680 Fax 1 862 209 4681 Mail orders na diagenode com PAGE 4 DIAGENODE AUTO IPure KIT v2 USER MANUAL Introduction The Diagenode IP Star Automated System automates immunoprecipitation and increases reproducibility Diagenode the leading provider of complete solutions for epigenetics research offers a variety of end to end systems to streamline DNA methylation and chromatin immunoprecipitation workflows Central to this full offering is Diagenode s Automated Systems simple yet robust automated bench top instruments that standardize different epigenetic applications i e ChIP MeDIP or MethylCap Diagenode designed these automation systems to make ChIP and DNA methylation studies accessible and reproducible and ensure consistent data in every experiment Diagenode Automated Systems will produce consistent results from any operator regardless of the day the experimental run or the lab Robust and reproducible results is a major goal of today s high resolution epigenomic studies Diagenode Automated Platforms replace the numerous manual error prone steps of complex epigenetic appl2. the Categories Name appears When selected the protocol on the protocol list the Run button shall turn executable Buttons e The user presses the Back button The user returns to the Protocols screen e The user presses the Shutdown button The screen shall be changed to Power Off e The user presses the Run button The screen shall be changed to Sample number e A Page up the list box e W Page down the list box Screen Sample number After the user presses the Run button the Sample number appears Buttons e he user presses the Sample number Text box The screen will be changed to keyboard e he user presses the Back button The user returns to the Protocol List screen e he user presses the Next button The screen shall be changed to Configuration or Layout information Europe Diagenode sa LIEGE SCIENCE PARK Rue Bois Saint Jean 3 4102 Seraing Ougr e Belgium Phone 432 4 364 20 50 Mail info diagenode com DIAGENODE AUTO IPure KIT v2 USER MANUAL PAGE 13 Sample number 7 E T DI Y 7 i i Keyboard diagenctie Layout information e Tip Rack DN70 Sample number 8 m Reagent information Laye 1 imfAvrmnantinn 700000000 0000000 2000000080 2 00000080 90000000 0000000 eaceccc ccQq co 00000080 000000000 0000000 400000060 00000000 t onawuor A Magnetic beads ul B
3. Belgium Phone 432 4 364 20 50 Mail info diagenode com DIAGENODE AUTO IPure KIT v2 USER MANUAL Troubleshooting Guide Error Cause Remedy SX 8G IP Star cannot be switched on SX 8G IP Star is not receiving power Check that the power cord is connected to the workstation and to the wall power outlet Computer cannot be switched on Computer is not receiving power Check that the power cord is connected to the computer and to the wall power outlet SX 8G IP Star shows no movement when a protocol is started SX 8G IP Star is not switched on Check that the SX 8G IP Star is switched on SX 8G IP Star shows abnormal movement when a protocol is started Aspirated liquid drips from the disposable tips Dripping is acceptable when ethanol is being handled For other liquids air is leaking The pipettor head may have lost its home position In the Software select Manual Operation Home After confirming that the pipettor head moves to the home position run the protocol again from the syringe pumps Grease or replace the O rings If the problem persists contact DIAGENODE Technical Services Technical Assistance At DIAGENODE we pride ourselves on the quality and availability of our technical support Our Technical Services Departments are staffed by experienced scientists with extensive practical and theoretical expertise in molecular biology and the use of DIAGENODE products If you have an
4. Total Action Time 0 Tithe no title Active Blacks 1 Tips Reagents Total Process time sec Complete Time 12 53 Prologue bat mx fle C Documents and Settingssl Mazon Desktop eichenhan protacolssM ew software proto time 0 7 he program will run through the following steps magnetic bead washes IP and IP washes During protocol the next window will be displayed indicating the current protocol step Europe Diagenode sa LIEGE SCIENCE PARK Rue Bois Saint Jean 3 4102 Seraing Ougr e Belgium Phone 32 4 364 20 50 Mail info diagenode com 3 5 Binding reaction 0 Start Watch Stack PUSH 40 Mix volume ul Repeat t Wait_msec 1000 waittime msec Stack DUP Liq in POP HP SPEED H Asp Speed Wait msec 200 wait time msec Stack DUP Liq out POP HP SPEED H Disp Speed Pass_time IF Goto LE 1800 Mixing Time Sec Repeat Stack Drop 9 The IP Star software indicates the end of the protocol Press the close buttom to finish the protocol run SX8G V52 Ver0 7 IPure 8 samples HLD 1 2 1 New Tip Collect 0 DIAGENODE AUTO IPure KIT v2 USER MANUAL y 0 1 1 2 Well information 0 1 1 3 E 0 1 3 1 Well 1 1 3 2 Bottom Sense 0 1 3 3 Pre asp 1 3 4 Beads resuspend 1 3 5 Binding reaction 1 3 6 Air Dsp 1 3 7 Interium Height Z Move 1 3 8 Tip Discard 1 4 1 New Tip C
5. permission of the authors Bioruptor and IP Star are registered trademarks of Diagenode
6. 16 16 rxns Auto Transcription ChIP kit protein A x100 C01010032 AB Auto03 A100 100 rxns Auto Transcription ChIP kit protein G x16 C01010031 AB Auto03 C016 16 rxns Auto Transcription ChIP kit protein G x100 C01010033 AB Auto03 G100 100 rxns Auto ChIP kit protein A x100 C01010011 AB Auto01 A100 100 rxns Auto ChIP kit protein G x100 C01010013 AB Auto01 G100 100 rxns Auto MeDIP kit x16 C02010011 AF Auto01 0016 16 rxns Auto MeDIP kit x100 C02010012 AF Auto01 0100 100 rxns Auto hMeDIP kit x16 C02010033 AF Auto02 0016 16 rxns Auto MethylCap x48 C02020011 AF Auto01 0048 48 rxns Auto IPure kit v2 C03010010 AL Auto01 0100 100 rxns Visit us at one of Diagenode s demo sites or discover our Automated Systems by performing some assays with the help of our R amp D and Technical Department Diagenode s a BELGIUM EUROPE LIEGE SCIENCE PARK Rue Bois Saint Jean 3 4102 Seraing Belgium Tel 32 4 364 20 50 Fax 32 4 364 20 51 orders diagenode com Diagenode Inc USA NORTH AMERICA 400 Morris Avenue Suite 101 Denville NJ 07834 USA Tel 1 862 209 4680 Fax 1 862 209 4681 orders na diagenode com For a complete listing of Diagenode s international distributors visit www diagenode com en company distributors php For rest of the world please contact Diagenode s a info na diagenode com info diagenode com 2014 Diagenode Inc All rights reserved The content of this document cannot be reproduced without prior
7. Manual ChIP Four different operators have each performed two ChIP experiments using H3K9me3 antibody on the genomic region SAT2 positive locus 10 000 Hela cells have been used per IP Reagents and sheared chromatin were identical per assay The standard deviations between the ChIPs performed by the same operator and between the four different operators are displayed Figure 2 Automated ChlP Four ChIP experiments using H3K9me3 antibody on the genomic region SAT2 positive locus have been performed by the SX 86 IP Star 10 000 Hela cells have been used per IP Reagents and sheared chromatin were identical per assay The standard deviations between the four ChIPs performed by the SX 8G IP Star are displayed PAGE 7 Diagenode Inc North America Phone 1 862 209 4680 Fax 1 862 209 4681 Mail orders na diagenode com PAGE 8 DIAGENODE AUTO IPure KIT v2 USER MANUAL Kit Method Overview Pure after ChIP Pure after MeDIP STEP 1 Chromatin reverse cross linking and elution Chromatin is decrosslinked and eluted from magnetic beads magnetic or agarose which are discarded Magnetic beads for purification are added STEP 2 DNA binding Magnetic beads acquire positive charge to bind negatively charged phosphate backbone of DNA DNA bead complex is separated using a magnet STEP 3 Washes Proteins and antibody debris are washed away STEP 4 DNA Elution DNA is eluted from magnetic beads w
8. P kit Auto Histone ChIP seq kit Auto Histone ChIP seq kit Auto MeDIP kit Auto MethylCap kit Auto hMeDIP Auto IPure kit v2 Reduces cross contamination Y Diagenode Inc North America Phone 1 862 209 4680 Fax 1 862 209 4681 Mail orders na diagenode com DIAGENODE AUTO IPure KIT v2 USER MANUAL Applications Software User interface User friendly Dispensing Protocol optimization flexible parameters New protocol development Characteristics ChIP seq MeDIP seq MethylCap seq hMeDIP IPure Sample preparation Re ChIP MagBisulfite RNA IP Library preparation for NGS platforms Protocols Sample prep MeDIP hMeDIP MethylCap e re EI Re ChIP MagBisulfite RNA IP e E a 7 7A y om IPure Library prep E diagencgde Intuitive touch screen panel Software training not required Automated dispension of assay reagents Antibody coating temperature time mixing speed Immunoprecipitation temperature time mixing speed Washes temperature time mixing speed Achievable by Diagenode product specialist 750W x 740 D x 610 H 100 kg 8 Nozzles X Y 7 axis 95 C ChIP seq MeDIP seq MethylCap seq hMeDIP IPure Sample preparation Re ChIP MagBisulfite RNA IP PC Software Software training before use Manual dispension of assay reagents Antibody coating temperature time Immunoprecipitation temperature time Achievable by customer a
9. diageng e Innovating Epigenetic Solutions AUTO IPure KIT v2 MANUAL Magnetic DNA Purification kit for epigenetic applications Auto IPure kit v2 x100 New Cat No C03010010 Old Cat No AL Auto01 0100 Version 41 19 09 14 Technical Assistance amp Ordering Information Diagenode s a BELGIUM EUROPE Diagenode Inc USA NORTH AMERICA LIEGE SCIENCE PARK 400 Morris Avenue Suite 101 Rue Bois Saint Jean 3 Denville NJ 07834 USA 4102 Seraing Belgium Tel 1 862 209 4680 Tel 32 4 364 20 50 Fax 1 862 209 4681 Fax 32 4 364 20 51 techsupport na diagenode com techsupport diagenode com orders na diagenode com orders diagenode com For a complete listing of Diagenode s international distributors visit http www diagenode com en company distributors php For the rest of the world please contact Diagenode s a DIAGENODE AUTO IPure KIT v2 USER MANUAL PAGE 3 Contents TO AOE OTe e pi a TUTTI TO QT QI TO TTL DO EE EA r E 4 IP Star Automated System for ChIP MeDIP amp MBD eeeeseee nr 5 Mit MemOd ONCRVICW C 8 dur i ee ee ee ee ee ee ee eee ee ee ee ees eee eee 9 TS OT N E E E EA E E EE E E EE E E 7 How to perform Automated IPure in the IP Star Compact ccc eee 10 PUG a con P PT 11 How to perform Automated IPure in the IP Star ce enhn 16 Oa CMG us running protocol PPP 17
10. dule in the workstation LEE o RR CEELI LE LAE OE E eee 53 8S ee tt E CEE E E E ee ew 233 oo oo oo s3ssssseses HMM amp bhes PARRA 4a PSATRE TA Lee ee eee ee STE a gi IIIITIIIAM easet L TRALEE acaumaeuaaqequSa Fi bees se ee amp a a a ae Poe eee hie eee ee TER E E E ee Steerer 44 hae ae a EREET EE ee Cs 12 11 10 Elution 2 Elution 1 Wash 2 Wash 1 Binding reaction Elution 2 Elution 1 Wash 2 Wash 1 Binding reaction Buffer C N CO A Oe YN CO Diagenode Inc North America Phone 1 862 209 4680 Fax 1 862 209 4681 Mail orders na diagenode com PAGE 20 DIAGENODE AUTO IPure KIT v2 USER MANUAL 5 Press the following icon a Select the protocol of interest Press start w Easy Protocol Start Screen Please make sure your selection IPure 18 samples McIP HLD IPure 16 samples HLD IPure 8 samples McIP HLD IPure 8 samples HLD IPure bisulfite Irina HLD IPure RNA Organic solvent HLD IPure RNA Standard HLD IPure cultured cell HLD IPure Whole blood HLD IPure PCR amplicon HLD MagPurification DINA from Bisulfite soution3 HLD 6 Before starting the protocol a start confirmation window will appear Press OK and the protocol will run Ca L1 p t Schedule Manager Ver1 0 Protocol CADocuments and Settings Mazon Desktop iweichenhan protacals New software a Min Sleep Timez 0 z i Total Sleep Time Max Action Times 0
11. fter training 1070W x 650 D x 780 H 130 kg 8 Nozzles X Y Z axis 4 95 C Europe Diagenode sa LIEGE SCIENCE PARK Rue Bois Saint Jean 3 4102 Seraing Ougr e Belgium Phone 32 4 364 20 50 Mail info diagenode com Improved reproducibility DIAGENODE AUTO IPure KIT v2 USER MANUAL Our SX 8G IP Star will increase the immunoprecipitation reproducibility between IPs performed by the same as well as by different operators see figure 1 and 2 below Reagents Antibodies buffers and sheared chromatin were identical for ManChIP and AutoChIP The SX 8G IP Star Automated system removes variation that can be created by manual handling and allows you to optimize and standardize your assay within a lab The SX 8G IP Star is designed to improve the accuracy and the reproducibility of any immunoprecipitiation experiment Man ChIP SD gG 0 9496 SD H3K9me3 11 3696 100 0 A 80 0 ChIP 1 ChIP 2 of Input 60 0 50 70 40 0 34 63 Auto ChIP 100 0 70 0 ChIP 1 56 25 of Input 1 26 IgG H3K9me3 SD IgG 0 69 SD H3K9me3 23 84 SD IgG 1 496 SD H3K9me3 2 3896 C SD IgG 0 1796 SD H3K9me3 1 1296 ChIP 1 ChIP 2 B SD IgG 0 0996 78 62 95 26 SD H3K9me3 0 65 D ChIP 1 ChIP 2 43 83 44 75 2 06 1 42 1 54 f 1 42 l SD IgG 0 28 SD H3K9me3 1 6 ChIP 1 ChIP 2 57 83 56 25 ChIP 2 ChIP 3 ChIP 4 54 71 57 83 54 34 1 00 1 45 0 81 IgG H3K9me3 IgG H3K9me3 IgG H3K9me3 Figure 1
12. hich are discarded Purified DNA is ready for any downs tream application qPCR next generation sequencing amplification microarray STEP 1 DNA Elution DNA is eluted from beads magnetic or agarose Magnetic beads for purification are 2 added STEP 2 DNA binding Magnetic Beads acquire positive charge e 5 to bind negatively charged phosphate backbone of DNA DNA bead complex Is separated by using a magnet STEP 3 Washes Proteins and antibody debris are x S 5 washed away STEP 4 DNA Elution DNA is eluted from magnetic beads which are discarded k gt Purified DNA is ready for any downs o tream application qPCR next generation sequencing amplification microarray Europe Diagenode sa LIEGE SCIENCE PARK Rue Bois Saint Jean 3 4102 Seraing Ougr e Belgium Phone 432 4 364 20 50 Mail info diagenode com DIAGENODE AUTO IPure KIT v2 USER MANUAL PAGE 9 Kit Materials Kit contents The kit content is sufficient to perform 100 reactions IPure kit 100 reactions Description Format Storage Wash buffer 1 w o iso propanol Wash buffer 2 w o iso propanol This product is shipped at 4 C Store it at 20 C upon arrival Plastics and consumables available separately Description Cat No Format 200 pl tube strips 12 tubes strip cap strips C30020001 80 dd dir strips 8 tubes strip cap strips for IP Star C30020002 120 96 well microp
13. ications with a reliable highly consistent and automated process that requires minimal operator intervention We empower researchers to simplify the tedious protocols and the complexity of many epigenetic protocols In addition Diagenode Automated Systems minimize sample carryover data variability and costly errors The platforms offer full workflow support for epigenetics research utilizing our complete kits and laboratory validated protocols to rapidly deliver high quality and consistent data Auto IPure kit v2 Diagenode s Auto IPure kit v2 is the only DNA purification kit using magnetic beads that is specifically optimized for extracting DNA from ChIP and MeDIP Chromatin IP and Methylated DNA IP experiments It s a simple and straightforward protocol that delivers pure DNA ready for any downstream application e g next generation sequencing This approach guarantees a minimal loss of DNA and reaches significantly higher yields than a column purification see results page Comparing to phenol chloroform extraction the IPure technology has the advantage of being nontoxic and much easier to be carried out on multiple samples The use of the magnetic beads allows for a clear separation of DNA and increases therefore the reproducibility of your DNA purification Diagenode s IP Star system uses the principle of bead based magnetic separation Magnetic beads bound with chromatin or DNA are brought to the inner wall of the tip when a strong
14. lates for IP Star C30080030 10 Tips box C30040021 960 Tips bulk C30040020 1000 2 ml microtube for IP Star Compact C30010014 100 Large reagent container for IP Star Compact C30020004 20 Medium reagent container for IP Star Compact C30020003 10 Kits and Modules available separately Description Reference Quantity Diagenode Inc North America Phone 1 862 209 4680 Fax 1 862 209 4681 Mail orders na diagenode com IP STAR COMPACT How to perform Auto Pure on the IP Star Compact DIAGENODE AUTO IPure KIT v2 USER MANUAL PAGE 11 How to perform Automated Pure on the IP Star Compact Auto IPure is done in 96 well plates placed in the room temperature modules of the IP Star Compact Each 96 well plate will have capacity to run 8 or16 IPure samples A ChIP and MeDIP Elution buffer To perform DNA purification with Auto IPure elution steps after Auto ChIP and Auto MeDIP must be done using the Elution Buffer provided in the Auto IPure kit v2 Elution buffer 1 rxns Buffer A 115 4 ul Buffer B 4 6 ul 120 ul volume is calculating with 2096 of excess 100 ul Elution Buffer per sample are needed per IP or input sample to perform the elution ChIP MeDIP Elution Buffer provided in the IPure kit will be used as indicated in the Auto ChIP and Auto MeDIP user manuals B Prepare Auto IPure kit buffers Add as indicated below the suggested isopropanol v
15. lsopropanol ul C Wash buffer 1 ul D Wash buffer 2 ul E Buffer C ul aiugenotle Block Regent Tip Rack MTP Sample number Sample and carrier in well 1 and 7 Block PCR Tube Diagenode Inc North America Phone 1 862 209 4680 Fax 1 862 209 4681 Mail orders na diagenode com Screen Layout Information After the user presses the next button from Sample number screen or Configuration screen the Layout Information screen appears Buttons The user presses the Back button The user returns to the previous screen The user presses the Next button The screen shall be changes to Set confirmation When the user presses a block that block is magnified on the work surface layout background The magnified view provides a better display of the correct method setup for that block on the work surface Based on the selected protocol the user follows the indications provided in the screens to set up correctly the different reagents and samples Mure al T Fast Middle Low Speed list menu PAGE 14 DIAGENODE AUTO IPure KIT v2 USER MANUAL Running status Protocol name Progress Bar Remaining time LT Fa Remaining time Left block diageng e Screen Running After the user presses the Run button in the Set confirmation screen the Running screen appears Buttons e he user presses the Stop button Then screen shall be changed to Stop Dialog S
16. magnetic force is applied This differs from other systems that collect the bound DNA on the bottom of a reaction well resulting in cleaner assays and less carryover Europe Diagenode sa LIEGE SCIENCE PARK Rue Bois Saint Jean 3 4102 Seraing Ougr e Belgium Phone 432 4 364 20 50 Mail info diagenode com DIAGENODE AUTO IPure KIT v2 USER MANUAL PAGE 5 IP Star and IP Star Compact Systems for automa tion of epigenetic applications Diagenode has developed two automated platforms IP Star and IP Star Compact designed to increase your lab s productivity efficiency and experimental reproducibility The two automated platforms are capable of processing up to 16 samples per cycle The automated systems processes sheared chromatin or DNA to deliver purified DNA ready for qPCR amplification microarray and sequencing analysis Both the IP Star and IP Star Compact have an easy to use open software that provides you with flexibility This allows you to create your personal protocol according to your specific needs Major benefits of Diagenode Automated Platforms IP Star Compact IP Star y 4 l High resolution ChIP seq and MeDIP seq profiles Automated library preparation for Next Generation sequencing Reduces hands on time to just 30 minutes Reduces variability between operators and labs Ideal for low sample starting amounts E E 5 E E 5 Compatible with Diagenode Kits Auto ChI
17. oO gt O D DU lt Z Q 0 IPure Competitor Q Competitor Z column column MeDIP assays were performed using the MagMeDIP kit cat mc magme 048 The immunoprecipitated samples were purified with the IPure technology and two competitor kits competitor Q and Z The purified DNA was eluted in 50 ul of water and quantified with a Nanodrop DNA recovery after purification of MeDIP samples using IPure technology MeDIP Auto Pure m meDNA m unDNA B TSH2B 80 m GAPDH 70 60 90 40 30 20 10 100 90 of input MeDIP 1 MeDIP 2 MeDIP 3 MeDIP 4 Methyl DNA IP results obtained with our Auto MeDIP Kit and after DNA purification using the Auto IPure kit v2 Methyl DNA IP assays were performed using DNA from U20S cells and the Auto MeDIP kit Diagenode After MeDIP the DNA was purified using the Auto IPure kit v2 Experiments were run in the IP Star following Diagenode s protocols The IP was performed by including the kit internal controls together with the human DNA sample The internal positive and negative DNA controls included in the IP assay are methylated DNA meDNA and unmethylated DNA unDNA As positive and negative control regions a non methylated region in the GAPDH promoter and the methylated region of TSH2B were tested Results showed 4 different AutoMeDIP Auto IPure experiments run in the IP Star automated system Europe Diagenode sa LIEGE SCIENCE PARK Rue Bois Saint Jean 3 4102 Seraing Ougr e
18. olalvolume Sm Diagenode Inc North America Phone 1 862 209 4680 Fax 1 862 209 4681 Mail orders na diagenode com PAGE 18 DIAGENODE AUTO IPure KIT v2 USER MANUAL C Dispense prepared reagents in 96 well plate hd Elution 2 Elution 1 n eeeeeeee v ee000000 vs 8 oo000000 Binding reaction 7 6 Elution 2 1o sampies 5 Elution 1 L G0 wanz 3 e e o o Wash 1 8 Samples 2 e e e Binding reaction 1 00 OQ OQ Bufferc m EL Well Description Volumes SSS Alternatively elution can be done in 150 ul followed by ethanol precipitation if final volume must be reduced CNN Buffer C to be dispensed by the IP Star 29 ul CAM 2 2 Sample can be IP sample or input Europe Diagenode sa LIEGE SCIENCE PARK Rue Bois Saint Jean 3 4102 Seraing Ougr e Belgium Phone 432 4 364 20 50 Mail info diagenode com DIAGENODE AUTO IPure KIT v2 USER MANUAL PAGE 19 D Loading and running protocol Be sure that the computer connected to the robot never switches to the standby modus standby modus has to be inactivated Standby of the computer will lead to the abort of the protocol Reagent Preparation 15 min 1 Switch on the SX 8G IP Star The power switch is on the right side of the instrument 2 Switch on the computer LJ 3 Start SX 8G6 V52 software through SX 8G V52 the following icon 4 Place the prepared 96 well plate in the indicated room temperature mo
19. ollect 1 5 1 Assay Plate vvell 0 1 5 2 Z position 1 5 5 Interium Height Z Move 1 B 1 Assay plate Vell4 1 6 2 Bottom Sense 0 1 5 3 Disp BufferA 1 6 4 Disp air 1 6 5 Interium Height Z Move 1 7 1 Assay Plate vvell 05 1 7 2 Z position 1 7 3 Asp 30ul 1 7 4 Z up 1 7 5 Interium Height Z Move 1 8 1 Assay plate Vell5 1 8 2 Bottom Sense 0 1 8 3 Disp BufferA 1 8 4 Disp air 1 8 5 Interium Height Z Move 1 8 6 Tip Discard The program ended successfully IPure 16 samples McIP HLD IPure 16 samples HLD IPure 8 samples McIP HLD IPure 8 samples HLD IPure bisulfite Irina HLD IPure RNA Organic solvent HLD IPure RNA Standard HLD IPure cultured cell HLD IPure Whole blood HLO IPure PCR amplicon HLD MagPurification DNA from Bisulfite soution3 HLD 10 Collect your purified DNA This is your DNA ready for qPCR Shutting down the IP Star 1 Click on File and press End to close the software correctly 2 Switch off the computer and its monitor 3 Switch off the IP Star Robot power switch on the right side Note Ensure that the door is closed PAGE 21 Diagenode Inc North America Phone 1 862 209 4680 Fax 1 862 209 4681 Mail orders na diagenode com PAGE 22 DIAGENODE AUTO IPure KIT v2 USER MANUAL Results Comparison of DNA recovery after purification with IPure technology and competitor kits D gt 100
20. olumes to the corresponding Auto IPure kit v2 buffers Wash Buffer 1 Wash buffer 1 100 rxns Wash buffer 1 w o iso propanol Teswume Hm Wash Buffer 2 Wash buffer 2 100 rxns Wash buffer 2 w o iso propanol Toalvoume Sm Running a protocol Diagenode Splash Screen A0 Automated After the software start up screen disappears the Diagenode splash Epigenetic Applications screen is displayed for several seconds and then disappears on Magtration Technology Genetein software ver 1 0 Copyright 2010 diagenctie Diagenode Inc North America Phone 1 862 209 4680 Fax 1 862 209 4681 Mail orders na diagenode com PAGE 12 DIAGENODE AUTO IPure KIT v2 USER MANUAL Protocols Maintenance Information diagenctie Protocols 4 L Sample prep hMeDIP E p dE 24 Re ChIP MagBisulfite S yes IPure Library prep Jus O diagencgde diagengte Sample number Sample number diagenctie Start Screen Top menu After the Digenode splash screen disappears the start screen is displayed This is the first active window it allows the user to enter into three different parts of the software USER ACTIONS Buttons e Protocols e Maintenance for technical service e Information Diagenode contact details Protocols screen All available protocols are displayed on this screen Screen Categories Name Protocol List aa aa After the user presses the Categories Namel button
21. tatus screen is preferred as a progress bar that moves across the screen as the step progresses Running status Screen Running status This screen gives informations about the current running step of the protocol The user can check trough this screen the passed and remaining time of the experiment PIR a emaining time Left block 3 fefe fee zc Screen Finish End When the protocol is complete a window appears telling user the run is over The screen behind this window should be the Startup screen When OK is pressed then the Startup screen appears and the user can immediately begin to remove their sample and prepare for the next run e Open the door after pushing the OK button e Recover the samples and remove the consumables At this point user is expected to be ready to press RUN Buttons diagenctie e he user presses the OK button Then screen shall be changed to Categories Name Protocol List Europe Diagenode sa LIEGE SCIENCE PARK Rue Bois Saint Jean 3 4102 Seraing Ougr e Belgium Phone 432 4 364 20 50 Mail info diagenode com DIAGENODE AUTO IPure KIT v2 USER MANUAL PAGE 15 CAUTION Screen Caution When the protocol finishes the user can return to the protocol list screen A or warm the Do you want to start again a protocol peltier block screen B to eliminate possible condensation in the block diagenctie CAUTION Temperature unit main
22. tenance is executed diagenctie diagenctie CAUTION Temperature unit maintenance Remaining time Left block gt Right block 2 diagenctie Diagenode Inc North America Phone 1 862 209 4680 Fax 1 862 209 4681 Mail orders na diagenode com How to perform Auto Pure on the IP Star IP STAR DIAGENODE AUTO IPure KIT v2 USER MANUAL PAGE 17 How to perform Automated Pure on the IP Star Auto IPure is done in 96 well plates placed in the room temperature modules of the IP Star Each 96 well plate will have capacity to run 8 or16 IPure samples A ChIP and MeDIP Elution buffer To perform DNA purification with Auto IPure elution steps after Auto ChIP and Auto MeDIP must be done using the Elution Buffer provided in the Auto IPure kit v2 Elution buffer 1 rxns Buffer A 115 4 ul Buffer B 4 6 ul volume is calculating with 2096 of excess 100 ul Elution Buffer per sample are needed per IP or input sample to perform the elution ChIP MeDIP Elution Buffer provided in the IPure kit will be used as indicated in the Auto ChIP and Auto MeDIP user manuals B Prepare Auto IPure kit v2 buffers Add as indicated below the suggested isopropanol volumes to the corresponding Auto IPure kit v2 buffers Wash Buffer 1 Wash buffer 1 100 rxns Wash buffer 1 w o iso propanol Iso propanol Total volume Wash Buffer 2 Wash buffer 2 100 rxns Wash buffer 2 w o iso propanol T
23. y questions or experience any difficulties regarding the SX 8G IP Star or DIAGENODE products in general do not hesitate to contact us DIAGENODE customers are a major source of information regarding advanced or specialized uses of our products This information is helpful to other scientists as well as to the researchers at DIAGENODE We therefore encourage you to contact us if you have any suggestions about product performance or new applications and techniques For technical assistance and more information call the DIAGENODE Technical Service Department or contact your local distributor PAGE 23 Diagenode Inc North America Phone 1 862 209 4680 Fax 1 862 209 4681 Mail orders na diagenode com Ordering information Description Cat No NEW Cat No OLD Format IP Star Compact B03000002 UH 002 0001 unit Auto True MicroChIP kit C01010140 16 rxns Auto True MicroChIP amp MicroPlex Library Prep Package C01010141 Poneman Te eee aa prep rxns MicroPlex Library Preparation kit x12 C05010010 AB 004 0012 12 rxns Auto Histone ChIP seg kit protein A x16 C01010020 AB Auto02 A016 16 rxns Auto Histone ChIP seg kit protein A x100 C01010022 AB Auto02 A100 100 rxns Auto Histone ChIP seg kit prowwtein G x16 C01010021 AB Auto02 G016 16 rxns Auto Histone ChIP seg kit protein G x100 C01010023 AB Auto02 G100 100 rxns Auto Transcription ChIP kit protein A x16 C01010030 AB Auto03 A0
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