This is a First Strand cDNA Synthesis Kit user manual, which contains all the reagents required for the reaction, and users only need to provide RNA samples for reverse transcription experiments.
The GeneCopoeia First Strand cDNA Synthesis Kit (C0210A) provides all reagents needed for reverse transcription of total RNA or poly(A) mRNA into single‑strand cDNA using M‑MLV Reverse Transcriptase (RNase H‑). The kit components include 5×RT Reaction Buffer, RNase Inhibitor, dNTPs, Oligo(dT)18, Random Primer, and Sequence‑Specific Primer, all supplied in a single‑use format.
The manual details storage conditions (‑20 °C), handling precautions, and a step‑by‑step protocol covering reagent preparation, reverse transcription (37 °C or 42 °C for 1 h depending on primer), enzyme inactivation (85 °C for 5 min), and downstream PCR usage without purification. An experimental example demonstrates validation of reverse transcription efficiency by GAPDH PCR. This manual is written in Chinese.
| Storage temperature | -20 °C |
|---|---|
| Total reaction volume | 13 µl |
| Oligo(dT)18 concentration | 60 µM |
| Random Primer concentration | 250 µM |
| M‑MLV Reverse Transcriptase activity | 200 U/µl |
The kit should be stored at –20 °C.
Incubate at 42 °C for 1 hour.
Incubate at 37 °C for 1 hour.
Yes, the first‑strand cDNA can be added directly to PCR reactions.