Genomic DNA Assay User Guide - PerkinElmer
Genomic DNAAssay User GuideFor LabChip GX Touch/GXII Touch Copyright 2014-2018, PerkinElmer, Inc. All rights reserved. PerkinElmer is a registeredtrademark of PerkinElmer, Inc. All other trademarks are the property of their respective owners.P/N CLS140166, Rev. DPublication Date: August 24, 2018
2ContentsSpecifications . 3Assay Specifications . 3Sample Conditions . 4Kit Contents . 4Safety and Usage . 6Safety Warnings and Precautions . 6Usage . 6Preparation Procedures . 7Additional Items Required . 7Preparing the Gel-Dye Solution . 7Preparing the DNA Samples, DNA Ladder and the Buffer Tube . 8Preparing the Chip . 9Inserting a Chip into the LabChip GX Touch/GXII Touch Instrument . 11Running the Assay . 13Cleaning and Storing the Chip . 16Chip Cartridge Cleaning . 17Results . 18Genomic DNA Software Analysis . 18Genomic DNA Ladder Result . 18Genomic DNA Result . 19Troubleshooting . 20LabChip Kit Essential Practices. 27General . 27Reagents . 28Chips . 29Samples . 32Chip Well Aspiration Using a Vacuum . 33Customer Technical Support . 34Licenses and Rights of Use . 35P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Specifications3SpecificationsAssay Specifications1Table 1. Assay SpecificationsSizing Range50 - 40,000 bpSizing Accuracy 20%Sizing Precision20% CVQuantitationRange0.2 - 5 ng/µLSensitivity0.1 ng/µLUp to 10 kb, based onladderFor water as samplebuffer. Final concentrationafter dilutionS/N 3; intact HumanControl gDNAQuantitationAccuracy 30%Based on PicoGreenquantitation of HumanControl gDNAQuantitationPrecision20% CVBased on Human ControlgDNASample VolumeRequired10 µL (minimum)20 µL(recommended)Requires a 384-well plate.A 96-well plate can beused but it requires aminimum of 20 µL and alow sip heightSamples perChip Prep48 or 24Two workflows: one for 24 samples, one for 48samplesAnalysis Time48 samples in 2.5hrsWalk-away timeSamples per480Chip Reagent KitChip Reagent Kit 3 - 9 monthsStability1.Human Control gDNA from intestine was purchased from BioChain (Hayward, CA).P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Specifications4Sample ConditionsTable 2. Sample ConditionsAdditivesPerkinElmer recommends that BSA anddetergents exceeding 0.05 mg/mL and0.01% v/v (respectively) in concentrationnot be used. Higher concentrations canresult in chip failure. In addition, nonaqueous solvents are not compatible withDNA LabChip protocols.ParticulatesAll sample plates should be spun downprior to analysis. All buffers should befiltered with a 0.22 µm cellulose acetatefilter.Salt ConcentrationTotal salt concentration must not exceed 10mM Tris. Higher salt concentrations anddifferent ions may alter performance andreduce assay sensitivity.Kit ContentsStorage: Store chips and reagents refrigerated at 2-8 C until nextuse. If using the chip again within 24 hours it may be left at roomtemperature. Allowing the chip wells to dry may lead to changes inchip performance.Kit contains enough reagents for 20 Small-batch or 10 Large-batchchip preparations. Up to 24 samples can be tested with a Smallbatch chip preparation. Up to 48 samples can be tested with aLarge-batch chip preparation.Table 3. Genomic DNA Reagent Kit Contents, PN CLS760685ReagentVialQuantityDNA Dye ConcentrateBlue1 vial, 0.09 mLDNA Chip StorageBufferWhite9 vials, 1.8 mL eachGenomic DNA GelMatrixRed5 vials, 1.1 mL each10X Genomic DNALadderYellow1 vial, 0.26 mLGenomic DNA MarkerGreen1 vial, 1.5 mLP/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Specifications5Table 4. Consumable ItemsItemSupplier and Catalog NumberQuantitySpin FiltersCostar, Cat. # 816010Detection Window VWR, Cat. # 21912-046Cleaning ClothSwab1ITW Texwipe , Cat. # TX758B3Table 5. DNA Extended Range LabChipItemCatalog NumberDNA Extended Range Chip (gDNA) for usewith GX Touch/GXII Touch HTCat. # 760517DNA Extended Range Chip (gDNA) for usewith GX Touch/GXII Touch 24Cat. # CLS138948P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Safety and Usage6Safety and UsageSafety Warnings and PrecautionsCAUTIONWe recommend that this product and components be handled onlyby those who have been trained in laboratory techniques and that itis used in accordance with the principles of good laboratorypractice. As all chemicals should be considered as potentiallyhazardous, it is advisable when handling chemical reagents to wearsuitable protective clothing, such as laboratory overalls, safetyglasses, and gloves. Care should be taken to avoid contact withskin or eyes. In case of contact with skin or eyes, wash immediatelywith waterWARNING!Dye Concentrate contains DMSO. S24/25: Avoid contact with skinand eyes.UsageThe Genomic DNA assay is for use with LabChip GX Touch/GXIITouch instruments. The LabChip GX Touch/GXII Touch instrumentsare for research use only and not for use in diagnostic procedures.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Preparation Procedures7Preparation ProceduresAdditional Items Required 18 megohm, 0.22-µm filtered water (Milli-Q or equivalent). 70% isopropanol solution in DI water. Bio-Rad Hard-Shell 384-well Skirted PCR Plates, Cat # HSP38XX (recommended). PerkinElmer Hard-Shell thin-wall 96-well skirted PCR plate(blue), Cat # 6008870 (recommended).Note: Allow the chip and reagents to equilibrate to roomtemperature at least 30 minutes before use.Preparing the Gel-Dye SolutionNotes: The Dye Solution contains DMSO and must be thawedcompletely before use.The dye is light sensitive. Do not expose the Dye solution or GelDye to light for any length of time. Keep the prepared Gel-Dyesolution in the dark.One vial of Genomic DNA Gel Matrix (red cap) is good for 4Small-batch or 2 Large-batch chip preparations. Up to 24 samplescan be tested with a Small-batch chip preparation. Up to 48samples can be tested with a Large-batch chip preparation.1Vortex the thawed Genomic DNA Dye Concentrate(blue cap) for 10 seconds before use.2Transfer 13.75 µL of Genomic DNA Dye Concentrate(blue cap) to 1 vial of Genomic DNA Gel Matrix (red cap).3Vortex the solution until it is well mixed and spin down for a fewseconds.4Transfer the mixture into two spin filters (approximately 550 µLeach).5Centrifuge at 9200 rcf for 7.5 minutes at room temperature.6Discard filters, label and date the tubes.7Store in the dark at 2-8 C. Use within 3 weeks.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Preparation Procedures8Preparing the DNA Samples, DNA Ladder and theBuffer TubeSample Workflow384-well Sample PlateSamples should be diluted in waterMaximum samples per run 48A 96-well plate can be used but it requires a minimum of20 µL and a low sip height. See Additional Items Requiredsection for plate-type compatibility.1X Ladder 12 µL DNA Ladder108 µL water Note: Do not vortex laddersolution. Vortexing may degradelarge DNA ladder fragmentsLadderTube750 µL waterBufferTubeLabChip GX Touch/GXII TouchFigure 1. Sample Workflow.1In the provided 0.2 mL Ladder Tube, add 12 µL of GenomicDNA Ladder (yellow cap) to 108 µL water (Milli-Q orequivalent). Mix thoroughly by pipetting the solution up anddown a few times. Avoid creating air bubbles. Ensure there areno air bubbles in the Ladder Tube.Note: Do not vortex the ladder solution. Vortexing may degradelarge DNA ladder fragments.2Insert the Ladder Tube into the ladder slot on the LabChip GXTouch/GXII Touch instrument.3Prepare samples in 384-well plates. Add 2 µL of sample to18 µL of water (Milli-Q or equivalent). If sample volume islimited, 1 µL of sample can be added to 9 µL of water.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Preparation Procedures9Notes: Due to evaporation, samples prepared with 1 µL of sampleand 9 µL of water should only be tested in Small-batch runs, 24samples.Samples at concentrations from 0.2 to 5 ng/µL should be runundiluted.Do not exceed 5 ng/µL final concentration in the well as this canclog the chip channels.We recommend testing intact DNA at 2.5 ng/µL4Add 750 µL of water (Milli-Q or equivalent) to the 0.75 mLBuffer Tube provided with the reagent kit. Ensure there are noair bubbles in the Buffer Tube.5Insert the Buffer Tube into the buffer slot on the LabChip GXTouch/GXII Touch instrument.Buffer TubeLadder TubeSampleWell A1LadderTubeBufferTubeFigure 2. Locations of the Buffer Tube and Ladder Tube in theGX Touch/GXII Touch instrument.Preparing the Chip1Allow the chip to equilibrate to room temperature at least 30minutes before use.2Use a pipette tip attached to a vacuum line to thoroughlyaspirate all fluid from the chip wells (see Figure 3). For moredetails on how to set up a vacuum line see page 33.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Preparation Procedures10Figure 3. Using a vacuum to aspirate the chip wells is moreeffective than using a pipette.3Rinse and completely aspirate each active chip well (1, 3, 4, 7,8, and 10) twice with water (Milli-Q or equivalent). Do not allowactive wells to remain dry.4If any water spills onto the top and bottom chip surfaces duringrinsing, aspirate using the vacuum line. DO NOT run the tip overthe central region of the detection window. Use the providedDetection Window Cleaning Cloth dampened in water (Milli-Q or equivalent) or alcohol to clean the chip detection window asneeded.5Using a reverse pipetting technique, add Gel-Dye solution tochip wells 3, 7, 8, and 10. For Small-batch, add 50 µL per wellas shown in Figure 4. For Large-batch, add 75 µL in wells 3, 7and 8 and 120 µL in well and 10 as shown in Figure 5.Figure 4. Reagent placementfor Small-batch (up to 24samples).P/N CLS140166, Rev. DFigure 5. Reagent placementfor Large-batch (up to 48samples).Genomic DNA Assay User GuidePerkinElmer, Inc.
Preparation Procedures611Add 60 µL Genomic DNA Marker (green cap) to chip well 4for Small-batch (Figure 4) or 120 µL for Large-batch (Figure 5).Note: The marker well may need to be replenished if the chip is inidle mode on the instrument for an extended period of time.7Make sure the rims of the chip wells are clean and dry.8IMPORTANT: Ensure chip well 1 (waste well) is empty beforeplacing the chip into the instrument.Inserting a Chip into the LabChip GX Touch/GXII TouchInstrument1Check that the sample plate, Buffer Tube, and Ladder Tube areplaced on the instrument properly.2Remove the chip from the chip storage container and inspectthe chip window. Clean BOTH sides of the chip window with thePerkinElmer-supplied clean-room cloth dampened with a 70%isopropanol solution in DI water.3Touch the Unload Chip button on the Home screen.Figure 6. Home screen.4Insert the chip into the LabChip GX Touch/GXII Touchinstrument (Figure 7) and close the chip door securely.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Preparation Procedures12Figure 7. Chip in the LabChip GX Touch/GXII Touch instrument.5Touch the Load Plate button on the Home screen (Figure 6) toretract the sample plate and send the sipper to the Buffer Tube.Note: Do not keep the chip door open for any length of time. Dye issensitive to light and can be photobleached.6The Assay Choice window will appear (Figure 8). Touch thedesired assay and then touch OK.Figure 8. Assay Choice menu.Notes: If performing multiple runs in a day, in between chippreparations the chip should be washed using the instrument andChip Storage buffer as described in “Cleaning and Storing the Chip”on page 16.Be sure to periodically clean the O-rings on the top plate of the chipinterface on the LabChip GX Touch/GXII Touch. Use the providedlint-free swab dampened with water to clean the O-rings using acircular motion. Allow the O-rings to dry before inserting a chip.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Preparation Procedures13Running the AssayNote: Chips can be primed independently from running assays.Touch the Prime button on the Home screen. Make sure the BufferTube is placed on the instrument.Figure 9. Chip priming screen.1Touch the Run button (see Figure 9).2Select the appropriate assay type (see Figure 8), plate name,well pattern, and whether to read wells in columns or rows.Select number of times each well is sampled under Adv.Settings (Figure 10). Touch the green arrow button.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Preparation Procedures14Figure 10. Selecting wells.3In the Setup Run tab, select the operator name, the option toread barcode, the destination of the file, the inclusion of samplenames, expected peaks, and excluded peaks and the filenameconvention. Select Auto Export to export results tablesautomatically (Figure 11). Touch the green arrow button.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Preparation Procedures15Figure 11. Run setup screen.4Touch Start to begin the run (Figure 12).Figure 12. Starting a run.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Preparation Procedures16Cleaning and Storing the ChipAfter use, the chip must be cleaned and stored in the chip container.1Place the chip into the plastic storage container. The sippershould be submerged in the fluid reservoir.2Remove the reagents from each well of the chip using vacuum.3Rinse and aspirate each active well (1, 3, 4, 7, 8, and 10) shouldtwice with water (Milli-Q or equivalent).4Add 100 µL of Storage Buffer (white cap5Place the chip in the LabChip GX Touch/GXII Touch instrument.Ensure that a Buffer Tube with 750 µL of water (Milli-Q orequivalent) is in the buffer slot.6Touch the Wash button in the upper right corner in the HomeScreen.The Wash screen opens (Figure 13).) to the active wells.Figure 13. Wash screen.7Remove the chip from the instrument and place it in the plasticstorage container.8Add an additional 50 µL of Storage Buffer to well 1.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Preparation Procedures917Cover the wells with Parafilm to prevent evaporation and storeat 2-8 C until next use. If using the chip again within 24 hours itmay be left at room temperature. Allowing the chip wells to drymay lead to changes in chip performance.Chip Cartridge Cleaning12DailyaInspect the inside of the chip cartridge and O-rings fordebris.bUse the provided lint-free swab dampened with water(Milli-Q or equivalent) to clean the O-rings using a circularmotion. If the O-rings stick to the chip or a pressure leak isdetected, perform the more extensive monthly cleaningprocedure.MonthlyaTo reduce pressure leaks at the chip interface, clean the Orings frequently. Remove the O-rings from the top plate ofthe chip interface on the LabChip GX Touch/GXII instrument.Soak O-rings in water (Milli-Q or equivalent) for a fewminutes. Clean the O-ring faces by rubbing between twofingers. Wear gloves.bTo reduce the occurrence of current leaks, clean the chipinterface frequently. Clean the top plate of the chip interfaceusing the provided lint free swab dampened with water (MilliQ or equivalent).cAllow the O-rings and chip interface to air dry. Reinsert theO-rings into the chip cartridge.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Results18ResultsGenomic DNA Software AnalysisData are analyzed by aligning the sample data and normalizing thesample area using the Lower Markers in the samples and in the twoladders that bracket every 12 samples. The size of a sample isdetermined by comparing the migration times of peaks within thesample to those of the fragments of known size in the bracketingladders. The concentration of a sample is calculated using a TotalgDNA smear, starting at 0.18 kb and extending to 300 kb (seeFigure 14). These smear limits can be adjusted by the user. Acalibration curve generated using the known size andconcentrations of ladder peaks are applied to the normalized areaof this Total gDNA smear, to determine the concentration of thesample. This Total gDNA Concentration is reported in the WellTable entry of each sample.The Genomic DNA assay also reports a Genomic DNA QualityScore (GQS) in the Well Table entry of each sample. This scorerepresents the degree of degradation of a sample, with 5corresponding to intact gDNA and 0 corresponding to highlydegraded gDNA, and is calculated from the size distribution of asample.Figure 14. Total gDNA smear (in orange).Genomic DNA Ladder ResultThe electropherogram of a typical Genomic DNA ladder is shown inFigure 15. Following the Lower Marker are ladder fragments of 100,300, 500, 700, 1100, 1900, 2900, 4900, 7000, 10000 and 40000 bp.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Results19LowerMarkerFigure 15. Genomic DNA Ladder.Genomic DNA ResultAn electropherogram and gel view of intact and degraded genomicDNA from BioChain is shown in Figure 16. Genomic DNA wasdegraded by incubating with Fragmentase for 10 or 20 minutes.Genomic DNA treated for 10 minutes was partially degraded(GQS 2.5), while genomic DNA treated for 20 minutes was highlydegraded (GQS 0).Figure 16. Intact and degraded genomic DNA from BioChain.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Troubleshooting20TroubleshootingNote: Some of the data examples shown in this section weregenerated with assays other than the assay described in this userguide.Symptom: Unexpected concentration and/or GQS.Possible causes and what to do:1If an unexpected concentration and/or GQS value is obtained,check the baseline of gDNA smear for a proper fit. Baselinescan be manually adjusted by clicking and dragging.Figure 17. Electropherograms of sample with a poor baselinefit and an adjusted baseline.Symptom: No ladder or sample peaks but marker peaks detected.Note: The lower marker peak height will most likely be greater thannormal height.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Troubleshooting21Possible causes:1Air bubble in sipper introduced during chip priming.What to do:1Reprime the chip. See “LabChip Kit Essential Practices” onpage 27 for instructions on how to reprime the chip.Symptom: Missing sample, ladder and marker peaks.Possible causes:1Clog in sipper or marker channel of chip.What to do:1Reprime the chip. See “LabChip Kit Essential Practices” onpage 27 for instructions on how to reprime the chip.Symptom: Ladder detected but no sample peaks.Possible causes:1The sipper is not reaching the sample due to low sample volumein the well of the plate.2If the missing sample peaks occurred only in a few wells of theplate, check those wells for air bubbles.3The sipper is not reaching the sample due to an incorrectcapillary height setting or incorrect plate definition.4If the plate has been uncovered for some time, sampleevaporation might have occurred.5Debris from the sample or sample prep is clogging the sipper.What to do:1Add more sample to the well.2Manually insert a larger volume pipette tip ( 100 µL) into thesample well and dislodge the bubble. Rerun these sample wells.3Check the plate definitions.4Check the sample wells, especially around the edge of the platewhere evaporation is fastest, and make a fresh plate if volumesare low.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Troubleshooting522If you suspect there may be debris in your samples, spin thesample plate down in a centrifuge (e.g. 3000 rcf for 5 minutes).Unclog the sipper by repriming the chip. See “LabChip KitEssential Practices” on page 27 for instructions on how toreprime the chip.Symptom: No ladder peaks but sample peaks and marker peaks arepresent.Possible causes:1Low or no ladder volume in the Ladder Tube.What to do:1Add more ladder to the Ladder Tube and restart the run.Recommended standard ladder volume is 120 µL (minimumvolume is 100 µL).Symptom: No marker peaks but sample peaks are present.Possible causes:1No marker added to chip well 4.2If there is marker solution in chip well 4, the problem may bedue to a marker channel clog.What to do:1This may be due to not filling marker well or chip remaining idleon instrument for extended period of time. Add or replenish themarker solution in the chip using the following procedure: Touch the Unload Chip button on the Home screen to openthe chip door. Return the chip to the chip container ensuring the sipper isimmersed in fluid. Thoroughly aspirate all fluid from chip well 4 using a vacuumline. Ensure that chip well 4 is rinsed and completely aspiratedtwice with water (Milli-Q or equivalent). Add Marker Solution (green cap) to chip well 4. Reinsert the chip back into the instrument. Restart the run.2Perform a marker channel unclogging procedure by reprimingthe chip. See “LabChip Kit Essential Practices” on page 27 forinstructions on how to reprime the chip.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Troubleshooting23Symptom: Ladder traces show up in the lanes following the ladders(delayed sip).Figure 18. Small ladder peaks in sample well caused bydelayed sip.Possible causes:1Separation channel overloaded with sample.2Partial clog in the separation channel.What to do:1Lower the starting sample concentration.2Reprime the chip. See “LabChip Kit Essential Practices” onpage 27 for instructions on how to reprime the chip.Symptom: Unexpected sharp peaks.Electropherograms of genomic DNA are shown in Figure 19 andFigure 20 with unexpected sharp peaks caused by particulates (likedust) or aggregates of large DNA that can form during sampleloading and/or migration in the chip.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Troubleshooting24Peak caused byparticulates orDNA aggregatesLowerMarkerFigure 19. Electropherogram with an unexpected peakmigrating after a genomic DNA sample.LowerMarkerPeak caused byparticulates orDNA aggregatesFigure 20. Electropherogram with an unexpected peak thatoverlaps with the migration of a genomic DNA sample.Possible causes:1Dust or other particulates introduced through sample orreagents.2Aggregates of large genomic DNA.What to do:1Replace the buffer used for sample and reagent preparation.Use a 0.22-micron filter for all water and buffers used for chip,sample, and reagent preparation.2Retest the sample. Dilute sample further, if possible, to reduceaggregation.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Troubleshooting25Symptom: Humps in several electropherograms which do notcorrespond to sample data.Figure 21. Humps in several electropherograms.Possible causes:1Electrode 7 is dirty and has contaminated the Gel-Dye mixturein well 7.What to do:1Before restarting the run, clean electrode 7. Remove the chipand follow the electrode cleaning procedure. We recommendusing the provided swab and isopropanol to manually cleanelectrode 7.Symptom: Peaks migrating much faster than expected.Note: Some migration time variance between chips or within a plateis considered normal chip performance. All chips are QC tested atPerkinElmer prior to shipmentPossible causes:1Incorrect Gel-Dye concentration.What to do:1Migration time is sensitive to dye concentration and peaks willmigrate too fast or too slow if the dye concentration in the gel istoo low or too high, respectively. Prepare a fresh Gel-Dyesolution. Wash and reprime the chip with the new Gel-Dyemixture.2If fast migration is observed repeatedly on a new chip, contactTechnical Support.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
Troubleshooting26Symptom: Peaks migrating much slower than expected.An electropherogram of a genomic DNA sample is shown inFigure 22 where the migration time is much longer than expected.LowerMarkerGenomic DNAFigure 22. Electropherogram of genomic DNA migration that istoo slow.Possible causes:1Particulates from the samples may be clogging the separationchannel.2Excess dye within the separation channel.3Gel-Dye was not primed properly into the chip.What to do:1Minimize the loading of particulates in the sample by spinningdown the plate (e.g. 3000 rcf for 5 minutes) before testingand/or selecting a plate type with a higher sip height in the StartRun dialog box before starting a new run. The debris maybeflushed out of the chip by washing and repriming the chip.2Prepare a fresh Gel-Dye solution. Wash the chip and thenreprime with the new Gel-Dye mixture.3Check the O-rings on the top surface of the chip interface andclean if necessary, then reprime the chip.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
LabChip Kit Essential Practices27LabChip Kit Essential PracticesTo ensure proper assay performance, please follow the importanthandling practices described below.Failure to observe theseguidelines may void the LabChip Kit product warranty.1Note: It is important to keep particulates out of the chip wells,channels and capillary. Many of the following guidelines aredesigned to keep the chips particulate-free.For assay and instrument troubleshooting, refer to the LabChip GXTouch software Help file or call PerkinElmer Technical Support at 1800-762-4000.General Allow the chip, sample plate and all reagents to equilibrate toroom temperature at least 30 minutes before use. Clean the O-rings in the chip interface weekly and theelectrodes daily. Refer to the Instrument Users GuideMaintenance and Service section for procedures. Avoid use of powdered gloves. Use only non-powdered gloveswhen handling chips, reagents, sample plates, and whencleaning the instrument electrodes and electrode block. Calibrate laboratory pipettes regularly to ensure proper reagentdispensing. Only the PerkinElmer-supplied clean room cloth can be used onthe chip to clean the detection window. Water used for chip preparation procedures must be18 megohm, 0.22-µm filtered water (Milli-Q or equivalent). Using the “Reverse Pipetting Technique” (described next) willhelp avoid introducing bubbles into the chip when pipetting thegel.1.PerkinElmer, Inc. warrants that the LabChip Kit meets specification at the time ofshipment, and is free from defects in material and workmanship. LabChip Kits arewarranted for 90 days from the date of shipment. All claims under this warrantymust be made within thirty days of the discovery of the defect.P/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
LabChip Kit Essential Practices28Reverse Pipetting TechniqueFigure 23. Reverse pipetting.1Depress the pipette plunger to the second stop.2Aspirate the selected volume plus an excess amount from thetube.3Dispense the selected volume into the corner of the well bydepressing plunger to the first stop.4Withdraw the pipette from the well. Store reagents at 2-8 C when not in use. The LabChip dye contains DMSO and should be thawedcompletely before use. It is recommended that you preparealiquots to reduce the time required for thawing. Gently vortex all kit reagents before use. Dispense reagents into chip wells slowly without introducing airbubbles. Insert the pipette tip vertically and to the bottom of thechip well. Protect the dye and Gel-Dye mixture from light. Store in the darkat 2-8 C when not in use. The Gel-Dye mixture expires 3 weeks after preparation.ReagentsP/N CLS140166, Rev. DGenomic DNA Assay User GuidePerkinElmer, Inc.
LabChip Kit Essential Practices29ChipsRepriming ChipsNote: Buffer tubes filled with 1X DNA sample buffer or water shouldbe placed into the instrument while priming or washing chips. Touch the Unload Chip button on the Home screen to open theinstrument door. Place the chip into the instrument. Close the chip door securely and choose the corresp
DNA Chip Storage Buffer White 9 vials, 1.8 mL each Genomic DNA Gel Matrix Red 5 vials, 1.1 mL each 10X Genomic DNA Ladder Yellow 1 vial, 0.26 mL Genomic DNA Marker Green 1 vial, 1.5 mL. Specifications 5 P/N CLS140166, Rev. D Genomic DNA Assay User Guide PerkinElmer, Inc. Table 4. Consumable Items
Magnetic beads for DNA purification 9 Genomic DNA purification kits 10 Genomic DNA extraction 16 Genotyping—pharmacogenomics studies 17 Plant genomic DNA isolation kits 18 Viral genomic DNA purification kits 20 Genomic DNA from saliva 21 Complete purification system for nucleic acids
approximately 60 -120 µg of total genomic DNA from haemolymph per isolate (50 µL) from the selected insects and the purity of genomic DNA ranged between 1.61 - 1.83 at 260 / 280 nm as revealed by spectrophotometry analysis. The quantity and quality of genomic DNA was compared with kit methods key. The electrophoretic analysis of the genomic
GENUS ABS JERSEY DIRECTORY Winter 2020 CONTENTS PROVEN/ GENOMIC SIRE NAME PAGE NO. PROVEN/ GENOMIC SIRE NAME PAGE NO. PROVEN/ GENOMIC SIRE NAME PAGE NO. Genomic CHEESEHEAD 3 Genomic LONESTAR 9 Proven VJ LARI 15 Proven COCHISE 4 Genomic MARIN
11. The DNA samples were stored at -20 C until further use. PCR amplification and gel electrophoresis PCR was carried out in a 50-μL reaction mixture, which contained 50 ng template DNA (CHO genomic DNA extracted as described above), 0. 25 U Taq DNA polymerase, 2.0 mM dNTPs, 1X Taq DNA polymerase buffer (Mg 2 plus) and 20 μM primer. The .
an ELISA TNF- assay to the DELFIA format. Conversion of other assays can be adapted from this example by referring to Table 3, which provides a specific recipe for the TNF- assay. 4 Table 2. Principles of ELISA and DELFIA TNF-α immunoassays ELISA DELFIA Assay schematic Assays An hTNF-ELISA assay was The hTNF-DELFIA assay used most of the
Genetic transformation and DNA DNA is the genetic material in bacterial viruses (phage) The base-pairing rule DNA structure. 2. Basis for polarity of SS DNA and anti-parallel complementary strands of DNA 3. DNA replication models 4. Mechanism of DNA replication: steps and molecular machinery
Recombinant DNA Technology 3. Recombinant DNA Technology 600 DNA ISOLATION AND PURIFICATION Basic to all biotechnology research is the ability to manipulate DNA. First and foremost for recombinant DNA work, researchers need a method to isolate DNA from different organisms. Isolating DNA from bacteria is the easiest procedure because bacterial cells
Year 5/6: Biographies – Joseph Briggs Lesson 1 Duration 1 hour. Date: Planned by Katrina Gray for Two Temple Place, 2014 Main teaching LO: To be able to recognise the features of a biography Cross curricular links: Literacy Q What is a biography? Link to the Greek prefix of ‘bio’ meaning ‘life’ Q What do you think are the features of a biography? Class teacher to make a list of pupil .
