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M-MLV Reverse Transcriptase

日期: 2015-05-26
瀏覽次數(shù): 234

貨號(hào):?28025013

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規(guī)格

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Optimal Reaction Temperature: 37° C
Final Product Size(s): 7 kb or less
Reverse Transcriptase: M-MLV Reverse Transcriptase
Ribonuclease H Activity: Reduced
Format: Tube(s)
Template: ssDNA,?ssRNA,?RNA-DNA Hybrid
Sensitivity: Low
Product Size: 40,000 units
Concentration: 200 U?μl
Enzyme Function: RNA dependent DNA Polymerase
Shipping Condition: Wet Ice
Number of Reactions: 200 Reactions

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內(nèi)容及儲(chǔ)存

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M-MLV RT is supplied with a vial of 5X first strand buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2 ], and a vial of 100 mM DTT. Store at -20°C. Guaranteed stable for 6 months when properly stored.

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描述

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M-MLV (Moloney Murine Leukemia Virus) Reverse Transcriptase (RT) is a recombinant DNA polymerase that synthesizes a complementary DNA strand from single-stranded RNA, DNA, or an RNA:DNA hybrid. Compared to AMV RT, M-MLV RT lacks DNA endonuclease activity and has a lower RNase H activity.

? Thermostability – Optimal activity at 37°C
? Size of cDNA – MMLV can be used to synthesize first-strand cDNA up to 7 kb
? Applications – Synthesis of first-strand cDNA (1), primer extension (2), sequencing dsDNA (3), cDNA libraries, and RT-PCR (4)

Source: Purified from E. coli expressing the pol gene of M-MLV on a plasmid (5).

Performance and Quality Testing: SDS-PAGE purity; endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; and yield and length of cDNA product.

Unit Definition: One unit of M-MLV RT is the amount of enzyme required to incorporate 1 nmole of deoxyribonucleotide into acid-precipitable material in 10 min. at 37°C using poly(A)?oligo(dT)12-18 as template?primer (6).

Unit Reaction Conditions: 50 mM Tris-HCl (pH 8.3), 40 mM KCl, 6 mM MgCl2, 1 mM DTT, 0.5 mM [3H]dTTP, 0.1 mM poly(A), 0.1 mM oligo(dT)12-18, 0.1 mg/ml BSA, and enzyme in 50 μl for 10 min. at 37°C (6).

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

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