HiScript ii Reverse Transcriptase
Introduction
HiScript II first-strand cDNA Synthesis kit Vazyme HiScript II reverse transcriptase is a new generation reverse transcriptase optimized from M-MLV (RNase H-) reverse transcriptase. The half-life of HiScript II at 50 ℃ is> 240 min. Even at 55 ℃, HiScript II can remain stable for a long time, which significantly benefits RNA transcription.
Templates with complex secondary structures. Furthermore, HiScript II has improved template affinity and improved cDNA synthesis efficiency. Has good resistance to most RT-PCR inhibitors and is suitable for amplification of long fragment cDNA (up to 20 kb).
The Vazyme HiScript II First Strand cDNA Synthesis Kit contains all the components necessary for first-strand cDNA synthesis. The products are suitable for PCR and qPCR. The 2 × RT Mix contains an optimized buffer and dNTP. HiScript II enzyme mix contains HiScript II reverse transcriptase and RNase inhibitor. Oligo- (dT) 23VN has a better affinity for Ploy A + RNA than Oligo (dT) 18. Additionally, random hexamers and gene-specific primers (GSP) are also optional.
Storage
All components should be stored at -20 ℃.
Protocol
Note:
1. Use high quality total RNA with high integrity for reverse transcription.
2. To avoid RNase contamination, keep the experiment area clean, wear clean gloves and masks, and use RNase-free tubes and tips.
3. Selection of primers (oligo- (dT) 23VN, random hexamers or GSP)
Whether the cDNA product will be used for PCR
- For eukaryotic RNA blots, generally use Oligo- (dT) 23VN to get the highest yield of full-length cDNA.
- Use a gene-specific primer (GSP) to obtain the highest specificity. However, switch to Oligo- (dT) 23VN or random hexamers if GSP fails on the 1 synthesis of st strand cDNA.
- Random hexamers with the lowest specificity for RNA templates, including mRNA, rRNA, and tRNA, can be used. Use random hexamers when Oligo- (dT) 23VN or GSP fail cDNA synthesis due to complex secondary structure, high GC content, or prokaryotic RNA template.
Whether the cDNA product will be used for qPCR
Use Oligo- (dT) 23VN mixture of random hexamers.
