A quick and standard operating guide for the Promega GoScript Reverse Transcription System (models A5000, A5001, A5003, A5004) to synthesize cDNA from…
This document provides a concise protocol for using the Promega GoScript Reverse Transcription System. It details two main procedures: a Quick Protocol suitable for general applications like Real-Time PCR, and a Standard Protocol optimized for longer mRNA or high GC content (>60%).
The guide covers essential optimization parameters including RNA template quality assessment, primer selection strategies (Oligo dT, Random Primers, Gene-specific), magnesium ion concentration adjustments, and reaction conditions such as denaturation temperature and extension time. This manual is written in Chinese.
| Reaction Volume (Quick) | 20 μl |
|---|---|
| RNA Template Amount (Quick) | 1-2 μg |
| Extension Time (Standard) | 60 min |
The suggested final magnesium ion concentration is 2.5 mM.
Yes, but the usage should not exceed one-fifth of the total reaction volume.
For total RNA, use 5 µg – 1 pg; for mRNA, use 500 ng – 100 fg.