GeneSpin Srl
Via Friuli, 51
20135 Milano
P.IVA C.F. 04520270960
Laboratory
and sales office
c/o Fondazione Filarete
Viale Ortles, 22/4
20139 Milano
tel. +39 02 56660199
fax. +39 02 537250
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 Leukemia Virus Reverse Transcriptase (M-MLV RT) lacks DNA endonuclease activity and has a lower RNase H activity.
STS-MRT10
10000 units
5X Reverse Buffer
STS-MRT50
50000 units
5X Reverse Buffer
component
STS-MRT10
STS-MRT50
M-MLV Reverse T.
10000 units / 50ul
50000 units / 250ul
5X Reverse Buffer
1ml 5X Buffer
1ml 5X Buffer
DTT
500ul / 100mM
500ul / 100mM
First Strand cDNA synthesis
Assay Set-Up:
Before starting, vortex all components thoroughly to ensure homogeneity.
Prepare a premix for the number of assays you need according to the following protocol:
First - Strand cDNA synthesis
Final volume: 20ul
- In a nuclease-free microcentrifuge tube add:
- dNTPs mix (10mM each) 1ul
- 1ng to 4ug total RNA
or
1-500ng mRNA xul
- 50-100ng random primers
or
Oligo(dT)12-18 (500ug/ml) 1ul
- ddH2O up to 12ul
- Heat mixture @ 65°C for 2min. and quick cool the tube on ice. Add:
- 5X Reverse Buffer 4ul
- 100mM DTT 2ul
- ddH2O 1ul
or
Ribonuclease Inhibitor
(optional)
- Mix gently and incubate
- @ 42°C for 2 min. if you are using oligo(dT)12-18
or
- @ 25°C for 2 min. if you are using random primers
- Add 1ul (200U) of GeneSpin M-MLV Reverse Transcriptase
(if you are using random primer incubate @25°C for 10 min.)
- Incubate @ 42°C for 60 - 120 min.
- Inactivate the reaction @ 70°C for 10 min.
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www.genespin.com
info@genespin.com
administration@pec.genespin.com
FOR RESEARCH USE ONLY
UNIT DEFINITION
One unit is defined as the amount of enzyme required to incorporate 10 nanomoles of dNTPs into acid-insolubile material in 30 min at 74°C.
SHIPPING
Shipped in green ice.
STORAGE
Store at -20C°
SHELF LIFE
12 months
FORM
Liquid
CONCENTRATION
200U/ul