M-MLV (H-) Reverse Transcriptase

概述:

Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) uses single-stranded RNA or DNA in the presence of a primer to synthesize a complementary DNA strand. It has a point mutation which leads to removal of RNase activity. Compared to other M-MLV mutants that only have the pol gene of M-MLV, M-MLV (H-) retains complete protein structure and polymerase activity. It can be used for synthesis of longer cDNA or preparation of cDNA library.

配方:

20 mM Tris-HCl, pH 7.5, 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol, 0.01% IGEPAL CA-630

参考文献:

1. Kimmel A R, et al: Preparation of cDNA and the generation of cDNA libraries: overview. Methods Enzymol. 1987, 152: 307-316.
2. Kievits T, et al: NASBA isothermal enzymatic in vitro nucleic acid amplification optimized for the diagnosis of HIV-1 infection. J Virol Methods. 1991, 35: 273-286.
3. Sun QM, et al: Expression, purification of humanVasostatin120-180 in Escherichia and its antiangiogenic characterization. Protein Expr Purif. 2005, 31: 311-317.

存储稳定性:

Store product at –20oC. For optimal performance aliquot product into smaller quantities after centrifugation and store at recommended temperature. Avoid repeated handling and multiple freeze/thaw cycles.

Components:

Component Name
M-MLV (H-) Reverse Transcriptase
5X RT Buffer

Specific Activity:

The specific activity of M-MLV (H-) Reverse Transcriptase was determined to be 200 U/µl as per the activity assay protocol, using the Unit Definition below,
Unit Definition:
One unit (U) is defined as the amount of enzyme that incorporates 1 nmol of dTTPs into acid-insoluble products in 10 min at 37°C with Poly (rA)-Oligo (dT) as the template/primer.

Applications of this product include: first strand synthesis of cDNA, in vitro reverse transcription, generation of cDNA libraries.