Manual

Product Number: RT03

Storage condition

-20°C

Component

MMLV (RNase H-) (200U/uL)

5×RT Buffer (with DTT)

Description

MMLV Reverse Transcriptase, encoded by Moloney Murine Leukemia Virus (MMLV RT) is an RNA-dependent DNA polymerase that synthesizes the complementary cDNA first strand from a single-stranded RNA template to which a primer has been hybridized. MMLV RT will also extend primers hybridized to single-stranded DNA. Second strand cDNA synthesis can be achieved from some mRNA templates without an additional DNA polymerase. The difference between this to the general MMLV RT is that the capacity to endure the heat is enhanced. It can remain the 100% activity at 50°C, it can also keep more than 80% activity even at 55°C.

Features

Lack RNase H activity: Weak RNase H activity High cDNA yield, can get more full-length cDNA.

Thermal stable: the optimum reaction temperature is 50°C, the highest is 55°C. Can overcome the template RNA secondary structure, and finish the reverse transcriptase experiment smoothly.

Wide temperature range: can reverse transcript from 37-55°C, with more than 80% of the highest activity at 42°C-55°C. customer can choose the reaction temperature freely.

Strong amplification activity: Gene mutation enhanced the binding capacity of the enzyme and RNA. Increased the amplification speed, can obtain the quality cDNA, suitable for cDNA library construction.

Source

Recombination of E. coli containing Moloney murine leukemia virus reverse transcriptase gene from clone of Moloney murine.

Application

The first-strand cDNA synthesis; RT-PCR.

Unit definition

         Using Poly (A) as the template and oligo (dT) as the primer, the enzyme required to catalyze the incorporation of 1 nmol of dTTP within 10 minutes at 37°C is defined as one active unit (U).

Storage Buffer

         20 mM Tris-HCl (pH7.5), 200 mM NaCl, 0.25 mM EDTA, 0.01% NP-40(v/v), 2.5 mM DTT, 50% glycerol (v/v).

5×Reaction Buffer:

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