1st Strand cDNA Synthesis Kit is a complete system for the efficient synthesis of first-strand cDNA. Reverse Transcriptase has dramatically improved thermal stability and can with stand reaction temperatures up to 50°C, making it suitable for reverse transcription of RNA templates with complex secondary structures. The Reverse Transcriptase also has an increased affinity for templates and is suitable for reverse transcription of small amounts of templates and low-copy genes. The Reverse Transcriptase also has an improved ability to synthesize full-length cDNAs, which can be amplified up to 10 kb in length.
The kit contains all the components needed to synthesize high-quality first-strand cDNA from total RNA or mRNA, and provides two primers for cDNA synthesis: Random Primers N6 and oligo(dT)18.The synthesized single-stranded cDNA product can be used directly in subsequent PCR or qPCR reactions.
| Reverse transcription reaction system preparation (20 μL system) | |
| Components | Volume |
| Total RNA | 1 ng -5 μg* |
| or mRNA | 1 ng-500 ng* |
| 5× Buffer | 4 μL |
| Random Primers N6 (50 μM) | 1 μL |
| or Oligo (dT)18 (50 μM) | or 1 μL |
| or Gene Specific Primers (2 μM) | or 1 μL |
| Enzyme Mix | 2 μL |
| RNase-free H2O | To 20 μL |
| Reaction program | |
| Reaction Temperature | Reaction Time |
| 25℃ | 5 min |
| 42℃ | 30 min |
| 85℃ | 5 min |