Applied Biosystems 7300 Real-Time PCR System (With RQ Study) SDS 1.3
I. ~ I. 1. : Dell GX280 2.8GHz with Dell 17 Flat monitor 256 MB RAM 40 GB hard drive DVD-RW drive Microsoft Windows XP Operating System with Service Pack 2 (SP2) 2. : (1) Sequence Detection Software V1.3 (2) Primer Express Software V3.0 (3) RQ Study Software 3. 7300 :
~ 7300 7300 New Document Wizard detectors Well Inspector plate well PCR Plate document (.sds ) data, Amplification Plot data baseline threshold data Spectra Component data
II. ~ 1. 7300 2. ABI Prism 7300 SDS 7300 icon : 3. : Disconnected Ready
File New Document Wizard Assay: Assay Applications When to use this Absolute Quantification For data collection and (Standard Curve) data analysis Relative Quantification ( for data collection Relative Quantification ( For data analysis Allelic Discrimination SNP genotyping For Post read PCR signal and SNP genotyping analysis Container: 96-well Clear Template: Blank Document
4. Next Detector Add Detector Plate Document Next Detector New Detector Detector Name: Reporter Dye: Quencher Dye: TaqMan Probe TAMRA MGB Probe none SYBR none Color: Detector Plate Document Detectors Create Another
5. Plate Document Wells Detector Use Task Unknown, NTC Standard ( Standard ) 6. Well Inspector Well Inspector ( ) :
7. Plate Document Template File menu Save As SDS Templates (*.sdt) 8. Template Plate Document File Save As SDS Document (*.sds) 9. PCR data ( )
Plate Document Instrument PCR Add Cycle, Add Hold, Add Step PCR : SYBR Add Dissociation Stage PCR Amplification Plot --- Results Amplification Plot Start PCR
10. running cycle run 1 Extend 1~20 cycle ( 3 cycle ) : Extend cycle Additional Comment Text
III. ~ 1. data Amplification Plot Threshold 2. Amplification Plot data Data menu Delta Rn vs Cycle Log Graph settings data
Analysis setting Auto-Ct baseline threshold Manual-Ct baseline threshold Auto-Ct Threshold Baseline Baseline Start End Cycles( End Cycle PCR 5 Cycle) Threshold: Log Threshold Geometric phase Baseline Threshold Analyze 3. Amplification Plot Spectra: filters raw data
Component: PCR Standard Curve: Standard Curve
Dissociation: SYBR Dissociation Curve non-specific non-specific
Report: data Report Excel Excel
IV. Relative Quantification Relative Quantification ( CT ) Relative Quantification ( CT ) Study 1. File New Document Assay Relative Quantification ( CT ) Study 2. OK Plate Document Add Plate RQ plate
3. single plate plate plate mutiplate document plate 10 plate document : a. PCR thermal cycle condition, sample volume data collection b. In use well sample name 4. analysis setting a. Threshold Baseline b. Gene Expression : Calibrator Sample Endogenous Control Detector
5. 6. Amplification Plot detector amplification plot
7. Gene Expression
V. ~ (8 Tubes/Strip)Optical Tubes (P/N4316567, 125 strips) (8 Caps/Strip)Optical Caps(P/N 4323032, 300 strips)
VI. : 1. ABI 7300 96 well optical plate (8 tubes/strip) optical tubes optical tube precision plate holder 2. PCR block 3. PCR block 4. run background plate update background file 5. : 0800251326 http://www.appliedbiosystems.com.tw