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Reverse Transcriptases

Home > PRODUCTS > Reverse Transcriptases

M-MLV Reverse Transcriptase

No Product Name Cat.# Conc Size Price(₩) Add
1 M-MLV Reverse Transcriptase RT001S 200 units/μl 10,000 units ₩ 80,000 추가
2 M-MLV Reverse Transcriptase RT001M 200 units/μl 20,000 units ₩ 144,000 추가
3 M-MLV Reverse Transcriptase RT001L 200 units/μl 50,000 units ₩ 320,000 추가
4 M-MLV Reverse Transcriptase RT001H 1,000 units/μl 50,000 units ₩ 320,000 추가
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Product description

Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) is a RNA-directed DNA polymerase that can synthesize a complementary DNA (cDNA) strand from a primer hybridized to either RNA or single stranded DNA as a template. M-MLV Reverse Transcriptase is capable of synthesizing DNA longer than 5 kb from messenger RNA. RNase H activity of M-MLV reverse transcriptase is weaker than that of Avian Myeloblastosis Virus (AMV) reverse transcriptase. 

 

Characteristics

- Molecular weight: 71 kDa

- Reaction temperature: 37 or 42

- Heat inactivation: 70, 10 min

 

Applications

- Synthesis of first-strand cDNA

- Primer extension

 

Quality control

- Purity: >99% on SDS-PAGE

- Endonuclease-free

- Exonuclease-free

- RNase-free

 

Unit definition

One unit is the amount of enzyme required to incorporate 1nmol of dTTP into acid-insoluble materials using 0.4 mM poly(rA)-oligo(dT) as substrate at 37 in 10 min.

 

Supplied with

- 10X M-MLV RT Buffer

- Sterile water (RNase free)

 

Storage buffer

50 mM Tris-HCl (pH 7.6), 150 mM NaCl, 1 mM DTT, 0.1 mM, EDTA, 0.1% NP40, 50% glycerol.

 

10X M-MLV RT Buffer

500 mM Tris-HCl (pH 8.3), 30 mM MgCl2, 100 mM DTT, 750 mM KCl

 

Standard reaction conditions

10X M-MLV RT Buffer

2 μl

M-MLV Reverse Transcriptase (200 units/μl)

1 μl

dNTP Mixture (2 mM each)

2 μl

a)Template RNA

X μl

b)Primer

1 μl

RNase Inhibitor (40 units/μl)

0.5 μl

Sterile water (RNase free)

up to 20 μl

a)Prepare one of the following RNA template.

    - Total RNA: 1 ng~5 μg

    - Messenger RNA (mRNA): 1 ng~250 ng

    - Specific RNA: 0.01 pg~0.5 μg

b)Prepare one of the following primers.

    - Oligo (dT)18: 50 μM~100 μM

    - Random hexamer: 50 μM~100 μM

    - Specific primer: 15 pmol~20 pmol

→An additional annealing step is recommended:

   - if using oligo(dT)18, incubate at 42 for 5 min.

   - if using random hexamer, incubate at 25 for 10 min.

→ Incubate at 37℃ or 42 for 60 min.

→ Incubate at 95 for 5 min to inactivate the reaction.

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