iScript™ IV Reverse Transcriptase

iScript™ IV Reverse Transcriptase (RT) is a novel MMLV mutant with superior robustness and reliability in RT reactions. It exhibits much higher efficiency in the first strand cDNA synthesis from RNA templates with secondary structures and high GC contents. The iScript™ IV Reverse Transcriptase is engineered to work under high temperatures (50°C-55°C), which can further facilitate to resolve the secondary structures and high GC problems of RNA. Besides, the iScript™ IV Reverse Transcriptase is significantly improved in inhibitor resistance, processivity, and reaction speed. iScript™ IV RT is designed to provide reliable, consistent, and fast cDNA synthesis in the presence of inhibitors found in a wide variety of samples.

The enzyme is available in 5,000 and 20,000 unit sizes at a concentration of 200 U/µL. The enzyme is supplied with a 5× Reaction Buffer.

Applications

• RT-PCR.
• Real Time RT-PCR.
• cDNA synthesis.

Product Specifications

• Storage Buffer: 20 mM Tris-HCl (pH 7.5 at 25^oC), 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.01% (v/v) NP-40, and 50% (v/v) glycerol.
•  Unit Definition: One unit of iScript™ IV RT is the amount of enzyme required to incorporate 1 nmole of dTTP into acid-precipitable material in 10 min at 37°C using poly(A)/ oligo(dT)₁₈ as a template/primer.
• Unit Reaction Conditions: 50 mM Tris-HCl (pH 8.3), 4 mM MgCl₂, 10 mM DTT, 50 mM KCl, 0.5 mM dTTP, 0.4 MBq/mL [³H]-dTTP, 0.4 mM poly(A)/oligo(dT)₁₈ and enzyme in 20 μl for 10 min at 37°C.