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.



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.

cat. no.

amount

note

PDF

STS-MRT10

10000 units

5X Reverse Buffer

STS-MRT50

50000 units

5X Reverse Buffer

STS-MRT200

200000 units

5X Reverse Buffer

Reverse Transcriptase

5X Master Mix

Primers

Oligonucleotides

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

@ 2008-2023 GeneSpin Srl

www.genespin.com

info@genespin.com

administration@pec.genespin.com

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