HelixCript™ Thermo Reverse Transcriptase, a thermostable and RNase H-negative variant of M-MLV RTase, can synthesize cDNAs from purified RNA templates at a temperature range of 42 ℃ ~ 55 ℃ and especially shows the highest activity at 50 ℃. At the escalated temperature, the cDNA synthesis is enhanced partially due to the less internal structural formation of the template RNA and increased polymerization activity of RTase. The high processivity and productivity of HelixCript™ Thermo Reverse Transcriptase allows this enzyme to amplify the high yield of product and can synthesize cDNA of target gene up to 12 kb from RNA template. This enzyme is quite suitable to synthesize the first-strand cDNA for RT-PCR of target gene. The cDNA synthesis from total RNA or poly-(A) RNA is performed by random primer, oligo-d(T) primer, or gene-specific primer.

 

 

Figure 1. The comparison between HelixCript™ Thermo Reverse Transcriptase and other brands of M-MLV RTase.

RT reactions, including 700 ng of human total RNA, were performed with each kit using reagents and conditions specified in each manufacturer’s guide. 10 % of each resulting cDNA was used in a PCR reaction containing 2.5 units of HelixAmp™ HyperSense-Taq DNA Polymerase and primer sets corresponding to each fragment of human dynein transcript. PCR products were analyzed by agarose gel electrophoresis.

Concentration

• Thermo Reverse Transcriptase : 200 units/μl

 

Storage

• -20 ± 5 ℃

 

Shelf life

• 12 months