Manual

Product Number: RT03GF

Shipping and Storage

Store at -20℃. Shelf‑life: 2 years.

Components

Description

Reverse transcriptase initiates synthesis of complementary DNA strand using RNA as template. Thermostable M‑MLV Reverse Transcriptase (RNase H) is a mutant variant of M‑MLV reverse transcriptase. Multiple point‑mutations eliminate the RNase H active site, which greatly diminishes RNase H activity. This reduces RNA degradation during reverse‑transcription reactions, improves yield of first‑strand cDNA and facilitates full‑length cDNA acquisition. Meanwhile, its thermal stability is enhanced, with optimal reaction temperature at 50℃.

Source

E. coli strain harbouring the gene for mutant Moloney Murine Leukemia Virus (M‑MLV) reverse transcriptase.

Unit Definition

One unit (U) is defined as the quantity of enzyme required to catalyze incorporation of 1nmol dTTP within 10 min at 37℃, using Poly(A) as template and Oligo(dT) as primer.

Application

First-strand cDNA synthesis; RT-PCR.

Storage Buffer

 20mM Tris-HCl (pH 7.5), 200mM NaCl, 0.25mM EDTA, 0.01 % NP-40 (v/v), 2.5mM DTT, 50 % Glycerol (v/v).

Purity Specification

After incubation of 200 U enzyme with 1μg 16S, 23S rRNA at 37℃ for 1 h, no visible change of electrophoretic bands can be observed.

Reaction Buffer

5× RT Buffer: 250mM Tris‑HCl (pH 8.3), 15mM MgCl₂, 375mM KCl, 50mM DTT.