This is the bulk version of HelixCript™ Thermo Reverse Transcriptase.

If you would like to purchase in bulk, please contact sale@nanohelix.net.

HelixCript™ Thermo Reverse Transcriptase, a thermostable and RNase H-negative variant of M-MLV reverse transcriptase (RTase), is an enzyme to synthesize cDNAs from RNA templates at a temperature range of 42 ℃ ~ 55 ℃ and shows the highest activity at 50 ℃. The high processivity and productivity of Thermo Reverse Transcriptase can synthesize cDNA up to 12 kb from an RNA template.

Application

• Generation of the first-strand cDNA up to 12 kb for libraries or cloning
• Two-step or one-step RT-PCR applications
• Conventional or real-time RT-PCR
• RT-PCR for detection of viral RNA

Figure 1. Comparison of activity of HelixCript™ Thermo Reverse Transcriptase with a competitor by RT-PCR.

Total RNAs were extracted from human blood. The first-strand cDNAs were synthesized using total 700 ng RNA, oligo-d(T) primer, 200 U HelixCript™ Thermo Reverse Transcriptase at 50 ℃ or 200 U Promega’s M-MLV RTase (Promega, USA) at 42 ℃. 10 % of cDNA reaction was used with a primer set corresponding to fragments of human dynein gene and 2.5 U HelixAmp™ HyperSense-Taq DNA Polymerase in 50 μl PCR reaction.

Enzyme activities: RNA-dependent DNA polymerase. No RNase H activity

Recommended reaction temperature: 42 to 65 ℃

Storage buffer: 20 mM Tris-HCl (pH 7.5), 100 mM KCl, 0.1 mM EDTA-NaOH (pH 8.0), 1 mM DTT, stabilizers, 50 % glycerol

Purity: ≥ 90 % (SDS-PAGE)

DNase contamination test: Not detectable (Incubation with 200 U enzyme and pUC19 plasmid at 37 ℃, 1 hr)

RNase contamination test: Not detectable (Incubation with 200 U enzyme and human total RNA at 37 ℃, 1 hr)

Activity test in cDNA synthesis, RT-PCR and qRT-PCR: Correspond to reference (Human RNA target)

Stability: 24 months at -20 ℃