ABI PRISM 7000 Sequence Detection Systems Relative ...

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Chapter 1 Introduction Materials and Equipment Materials and Equipment You need to supply the following items to complete an RQ study. Item High Capacity cDNA Archive Kit TaqMan ® Universal PCR Master Mix MicroAmp ® Optical 96-Well Reaction Plate and Optical Caps Note: Reaction plates and caps may also be purchased separately. Reaction plates can also be sealed with ABI PRISM Optical Adhesive Covers. Labeled primers and probes from one of the following sources: • Assays-on-Demand Gene Expression Products (predesigned primers and probes) • Assays-by-Design service (predesigned primers and probes) • Primer Express Software (customdesigned primers and probes) Applied Biosystems (PN 4322171) Applied Biosystems (PN 4304437) Applied Biosystems (PN 403012) Source • Applied Biosystems Web site • Contact your Applied Biosystems Sales Representative • PN 4330710 (1-user license) PN 4330709 (10-user license) PN 4330708 (50-user license) Reagent tubes with caps, 10-mL Centrifuge with adapter for 96-well plates Gloves Microcentrifuge Microcentrifuge tubes, sterile 1.5-mL Nuclease-free water Pipette tips, with filter plugs Pipettors, positive-displacement Tris-EDTA (TE) Buffer, pH 8.0 Vortexer Applied Biosystems (PN 4305932) Major laboratory supplier (MLS) Major laboratory supplier (MLS) MLS MLS MLS MLS MLS MLS MLS DRAFT September 26, 2003 3:14 pm, C1_Intro.fm Notes 4 Relative Quantification Getting Started Guide for 7000 v1.1
Materials and Equipment Description of the Sample RQ Experiment 1 DRAFT September 26, 2003 3:14 pm, C1_Intro.fm Notes Relative Quantification Getting Started Guide for 7000 v1.1 5
Page 1 and 2: 5′ cDNA Reverse Primer Oligo d(T)
Page 3 and 4: Primer Extended on mRNA 5′ 3′ 5
Page 5 and 6: Chapter 5 Performing an RQ Study 33
Page 7 and 8: Preface How to Use This Guide Purpo
Page 9 and 10: Safety and EMC Compliance Informati
Page 11 and 12: Symbols on Instruments Electrical S
Page 13 and 14: Chemical Safety Chemical Hazard War
Page 15 and 16: Physical Hazard Safety Ultraviolet
Page 17 and 18: Safety and Electromagnetic Compatib
Page 19 and 20: Introduction 1 About the 7000 SDS I
Page 21: About the Sample RQ Experiment Desc
Page 27 and 28: Specifying the Components of an RQ
Page 29 and 30: Selecting the Sequence Detection Ch
Page 31 and 32: Choosing the Probes and Primers Abo
Page 35 and 36: 5′ 3′ 5′ cDNA Reverse Primer
Page 39 and 40: Preparing the PCR Master Mix Preven
Page 41 and 42: Creating an RQ Plate Document Detec
Page 43 and 44: Specifying Thermal Cycling Conditio
Page 45 and 46: Analyzing and Viewing RQ Plate Data
Page 47 and 48: Analyzing and Viewing RQ Plate Data
Page 49 and 50: Exporting RQ Plate Documents Reanal
Page 53 and 54: Creating an RQ Study Document 5. Co
Page 55 and 56: Analyzing and Viewing the Results o
Page 57 and 58: Analyzing and Viewing the Results o
Page 59 and 60: Omitting Samples from a Study Omitt
Page 61 and 62: Exporting RQ Study Results Exportin
Page 63 and 64: References Kwok, S. and Higuchi, R.
Page 65 and 66: Creating Detectors A Before you can
Page 67 and 68: Upgrading the 7000 Software with RQ
Page 69 and 70: Index SAMPLE DOCUMENT September 26,
Page 71 and 72: Index SAMPLE DOCUMENT September 26,
Page 73 and 74:
DRAFT September 26, 2003 3:17 pm, G
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