Applied Biosystems 7900HT Fast Real-Time PCR System and SDS ...

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7900HT FAST Real-Time PCR SystemChapter 1Product OverviewAutomated OperationIn this mode, the Applied Biosystems 7900HT Fast Real-Time PCR System uses anAutomation Accessory (Zymark Twister Microplate Handler and fixed-position barcode reader), to provide high-throughput operation suitable for small- tomedium-scale studies. With this configuration (Figure 1-12), a technician can runbatches of plates unattended using the Automation Controller Software. As inStand-Alone operation, the computer stores all data for operation of the instrumentand provides the software for analyzing the run data.ComputerSDS SoftwareAutomation ControllerSoftware1Plate DocumentsCreatedGR2009GR20097900HTFront view with RobotApplied Biosystems 7900HTFast Real-Time PCR SystemInstrument Firmware3Thermal Cycling andSequence Detection25Plate document filesadded to and run fromthe plate queueRaw Data Saved tothe Plate DocumentsHard drive(s)*.sdsfiles*.sdtfile4Data CollectionSerialCable6Plate DocumentOpened and AnalyzedFigure 1-12Automated Operation of the of the 7900HT InstrumentDatabase OperationFor this mode, the Applied Biosystems 7900HT Fast Real-Time PCR System usesthe SDS Enterprise Database and the Automation Accessory, to provide maximumhigh-throughput potential. In this configuration (Figure 1-13 on page 1-19), the7900HT instrument has the storage capacity and analysis capability to supportmedium- to large-scale gene expression and genotyping studies. Moreover, the SDSEnterprise Database functions as a repository for all SDS-related data produced bythe networked client computers used for data collection and analysis.1-18 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User GuideDRAFTSeptember 1, 2004 11:38 am, CH_Overview.fm
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Page 1 and 2: Applied Biosystems7900HT Fast Real-
Page 3 and 4: ContentsPrefaceHow to Use This Guid
Page 5 and 6: Chapter 4Operating the InstrumentNo
Page 7 and 8: Chapter 7Maintaining the Instrument
Page 9 and 10: Appendix D Theory of OperationFluor
Page 11 and 12: PrefaceHow to Use This GuidePurpose
Page 13 and 14: Safety and EMC Compliance Informati
Page 15 and 16: Symbols on InstrumentsSymbols on In
Page 17 and 18: General Instrument SafetyGeneral In
Page 19 and 20: Chemical Waste SafetyChemical Safet
Page 21 and 22: Physical Hazard SafetyOvervoltageRa
Page 23 and 24: Workstation SafetyWorkstation Safet
Page 25 and 26: Product Overview 11In This Chapter
Page 27 and 28: 7900HT FAST Real-Time PCR SystemSec
Page 29 and 30: 7900HT InstrumentInterchangeableThe
Page 31 and 32: 7900HT FAST Real-Time PCR SystemBar
Page 33 and 34: Compatible ConsumablesCompatible Co
Page 35 and 36: Instrument ConnectionsFixed-Positio
Page 37 and 38: Section 1.2 Getting to Know the Sof
Page 39 and 40: SDS Software Related Files and Form
Page 41: 7900HT FAST Real-Time PCR SystemMan
Page 45 and 46: Section 1.3 SDS Enterprise Database
Page 47 and 48: * + Enter* +Enter3Down UpNumLockSys
Page 49 and 50: About the SDS Enterprise Database S
Page 51 and 52: Esc F1 F2 F3 F4 F5 F6 F7 F8 F9 F10
Page 53 and 54: Getting Started 22In This Chapter G
Page 55 and 56: Getting StartedUsing the SDSSoftwar
Page 57 and 58: GR23967900HTPowering On the 7900HT
Page 59 and 60: Using the SDS SoftwareUsing the SDS
Page 61 and 62: Using the SDS SoftwareAbout theMess
Page 63 and 64: Basic Software Skills TutorialNotes
Page 65 and 66: Basic Software Skills TutorialExerc
Page 67 and 68: Basic Software Skills TutorialExpor
Page 73 and 74: Basic Software Skills TutorialLesso
Page 77 and 78: Basic Software Skills Tutorial3. Se
Page 81 and 82: Using SDS Plate DocumentsUsing SDS
Page 83 and 84: Using SDS Plate DocumentsWell Inspe
Page 85 and 86: Preparing a Run 33In This Chapter N
Page 87 and 88: Notes for Database UsersDescription
Page 89 and 90: Workflow OverviewWorkflow OverviewE
Page 91 and 92: Workflow OverviewAllelicDiscriminat
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Step 1 - Creating a Plate DocumentS
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Step 2 - Applying Detectors and Mar
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Step 3 - Configuring the Plate Docu
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Step 5 - Programming the Plate Docu
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Step 6 - Saving the Plate Document
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Step 7 - Creating a Plate Document
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Step 9 - Running the Plate on the 7
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Operating the Instrument 44In This
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Notes for Database UsersDescription
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Section 4.1 Consumable PreparationS
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Compatible ConsumablesTable 4-1Comp
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Preparing Optical Plates for Usea.
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Preparing TaqMan Low Density Arrays
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Page 133 and 134:
Page 135 and 136:
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Section 4.2 Running an Individual P
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Running a Single Plate (Using the S
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Running a Single Plate (Using the S
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After the RunAfter the RunUser Acce
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Section 4.3 Automated OperationSect
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Operating the Software with an SDS
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Operating the Software without an S
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Page 153 and 154:
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Running Plates Using the Automation
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Running Plates Using the Automation
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Analyzing End-Point Data 55In This
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Notes for Database UsersNotes for D
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Section 5.1 Allelic DiscriminationS
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OverviewTable 5-1Signal and Sequenc
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Before You BeginBefore You BeginUsi
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Opening the Run DataOpening the Run
Page 171 and 172:
Calling and Scrutinizing Allelic Di
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Page 175 and 176:
Page 177 and 178:
After the AnalysisSaving theResults
Page 179 and 180:
Analyzing Real-Time Data 66In This
Page 181 and 182:
Page 183 and 184:
Section 6.1 Absolute Quantification
Page 185 and 186:
OverviewAlgorithmicManipulation ofR
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Analysis ChecklistAnalysis Checklis
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Analyzing the DataAnalyzing the Dat
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Viewing ResultsViewing ResultsViewi
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Section 6.2 Relative Quantification
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OverviewAlgorithmic Manipulation of
Page 197 and 198:
OverviewAutomatic Outlier RemovalAb
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Getting StartedGetting StartedUsing
Page 201 and 202:
Options for Analyzing Relative Quan
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Creating the Studyd. In the Plate Q
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Analyzing the Study• Manual Ct -
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Analyzing the StudySpecifying the A
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Viewing ResultsViewing ResultsDispl
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Viewing ResultsAbout DownArrowsDown
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After the AnalysisAfter the Analysi
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Section 6.3 Dissociation Curve Anal
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Before You BeginExample ResultsFigu
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Opening the Run DataOpening the Run
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Determining T m Values for the Anal
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Section 6.4 Procedure ReferenceSect
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Setting the Baseline and Threshold
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Eliminating Outlying AmplificationE
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Eliminating Outlying Amplification6
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After the AnalysisSaving theResults
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Maintaining the Instrument 77In Thi
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Section 7.1 Maintaining the 7900HT
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Replacing the Sample BlockMaterials
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Replacing the Sample Block9. Loosen
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7900HT FAST Real-Time PCR SystemRep
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Changing the Plate Adapter4. Place
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Decontaminating the Sample BlockCle
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Performing a Background RunPreparin
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Performing a Background RunAnalyzin
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Performing a Pure Dye RunAfter a ru
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Performing a Pure Dye Run• Fast 9
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Performing a Pure Dye RunAnalyzing
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Adding Custom Dyes to the Pure Dye
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Adding Custom Dyes to the Pure Dye
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Verifying Instrument PerformanceTab
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Verifying Instrument PerformancePre
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Section 7.2 Maintaining the Plate H
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Adjusting the Sensitivity of the Pl
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Adjusting the Sensitivity of the Pl
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7900HT FAST Real-Time PCR SystemAli
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Aligning the Plate Handler5. Using
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Aligning the Plate Handler7. Using
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Aligning the Plate Handler13. Click
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7900HT FAST Real-Time PCR SystemAli
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Aligning the Fixed-Position Bar Cod
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Section 7.3 Maintaining the Compute
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Maintaining the SDS softwareSeveral
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Troubleshooting 88In This Chapter T
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Troubleshooting TableTable 8-1Troub
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Low Precision or IrreproducibilityL
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Low Precision or IrreproducibilityD
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Background RunsBackground RunsBackg
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Pure Dye Runs6. Repeat step 4 until
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End-Point Runs (Allelic Discriminat
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Software and 7900HT InstrumentTable
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Zymark Twister Microplate Handler a
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TaqMan Low Density ArrayTable 8-6Tr
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Software ReferenceAAIn This Appendi
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Importing Plate Document Setup Tabl
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Setup Table Files1. File VersionDes
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Setup Table Files6. Detectors ListD
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Setup Table Files9. Markers List (A
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Setup Table FilesFormat(for a singl
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Setup Table FilesFormat(for a singl
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Setup Table FilesExample Code A-16A
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Exporting Plate Document Data4. In
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Operating the SDS Software from a C
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Using the Search ToolLogical Operat
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Connecting SDS Software to the Data
Page 333 and 334:
Designing TaqMan Reagent-BasedAssay
Page 335 and 336:
Assay Development Guidelines• For
Page 337 and 338:
Design Tips for Allelic Discriminat
Page 339 and 340:
Kits, Reagents, and ConsumablesCCIn
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Consumables and DisposablesConsumab
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Instrument Maintenance and Verifica
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Custom Oligonucleotide SynthesisTab
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Theory of OperationDDIn This Append
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Fluorescent-Based ChemistriesBasics
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Real-Time Data AnalysisPhase 2: Lin
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Real-Time Data AnalysisSignificance
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A similar equation for the endogeno
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Limited Warranty StatementEEWarrant
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BibliographyGeneral Paperson TaqMan
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Klein, D., Janda, P., Steinborn, R.
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Dolken, L., F. , Schuler, and G. ,
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AllelicDiscriminationVirtanen M., H
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Glossary.sdd.sdm.sds.sdt∆C T∆
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ackgroundbaselinecalibratorcDNA(com
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minor groovebinder (MGB)multicompon
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R nrunsessionsingle nucleotidepolym
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IndexSymbols- 6-31↑ 6-33Numerics7
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fluorescent, common sources 8-7isol
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adjusting step parameters 3-21confi
Page 381 and 382:
sample block locking bar 7-8sample
Page 384:
Worldwide Sales and SupportApplied
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