M-MLV(H-) Reverse Transcriptase
M-MLV reverse transcriptase 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, Moloney murine leukaemia virus reverse transcriptase (M-MLV RT) lacks DNA endonuclease activity and has lower RNase H activity.
Characteristics of this enzyme:
• Thermostability: optimal activity at 37 ° C
• cDNA size: M-MLV can be used to synthesize first-strand cDNA up to 7 kb
• Applications: first-strand cDNA synthesis, primer extension, dsDNA sequencing, cDNA libraries and RT-PCR
Source
Purified from E. coli expressing the M-MLV pol gene in a plasmid
Performance and quality testing
Purity SDS-PAGE; endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; and yield and length of cDNA product
Definition of unit
One unit of M-MLV RT is the amount of enzyme required to incorporate 1 nmol deoxyribonucleotide into acid precipitable material in 10 min at 37 ° C using poly (A) oligo (dT) 25 as template-primer.
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) 25 mM, 0.1 mg / ml BSA, and enzyme in 50 µL for 10 min at 37 ° C. For research use only. It should not be used in diagnostic procedures.
Specifications
Enzymatic function
RNA-dependent DNA polymerase
It includes
1 × 200 μL M-MLV RT (200U / μL); 1 ml of 5X first strand buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM magnesium chloride]; 500 µL 100mM DTT; 40,000 units
No. Of tests
200 reactions
Optimal reaction temperature: 37 ° C
Kind of product: M-MLV reverse transcriptase
Quantity: 200 reactions; 40,000 units
Reverse transcriptase: M-MLV
Ribonuclease H activity: Reduced
Sensitivity: Low
Shipping condition: Wet ice
Enough for 200 reactions
Template: ssDNA, ssRNA, hybrid RNA-DNA
Concentration: 200 U⁄µl
Format: Tube (s)
GC-rich PCR performance: Low
No. of reactions: 200 reactions, 200 reactions
PCR method: RT-PCR
Size (final product): 7 kb or less
Volume: 200 μl
Content and storage
• 1 × 200 µl M-MLV RT (200 U / µl)
• 1 ml of 5X first strand buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM magnesium chloride]
• 500 µl of 100 mM DTT
Store at -20 ° C. Stability guaranteed for 6 months when properly stored.
