GoScript Reverse Transcription System Protocol....英文文献资料.pdfVIP

T e c h n i c a l M a n u a l GoScript™ Reverse Transcription System INSTRUCTIONS FOR USE OF PRODUCTS A5000 AND A5001. PRINTED IN USA. Revised 12/12 Part# TM316 GoScript™ Reverse Transcription System All technical literature is available on the Internet at: /protocols/ Please visit the web site to verify that you are using the most current version of this Technical Manual. Please contact Promega Technical Services if you have questions on use of this system. E-mail: techserv@ 1. Description 1 2. Product Components and Storage Conditions 2 3. General Laboratory Precautions 3 4. Detailed Protocol 6 A. First-Strand cDNA Synthesis7 B. cDNA Quantification Using qPCR 8 C. PCR Amplification for Endpoint Analysis 9 D. Analysis11 5. Protocol Optimization11 A. GoScript™ Reverse Transcription System Reaction Characteristics11 B. RNA Template 12 C. Magnesium Concentration Optimization12 D. Primer Options and Design 13 E. Reverse Transcriptase Enzyme Concentration 13 F. Control Reactions 13 G. Temperature 14 6. Troubleshooting 15 7. References 20 8. Related Products 20 1. Description The GoScript™ Reverse Transcription System(a,b) is a convenient kit that includes a reverse transcriptase and an optimized set of reagents designed for efficient synthesis of first-strand cDNA optimized in preparation for PCR amplification. The components of the GoScript™ Reverse Transcription System can be used to reverse transcribe RNA templates starting with total RNA, poly(A)+ mRNA or synthetic transcript RNA. Figure 1 provides an overview of the reverse transcription proce