GENTAUR Contact Gentaur

Up

GENTAUR Survey

GENTAUR EUROPE

+32 1658 9045
+32 1650 9045

BELGIUM1

tel +32 2 732 5688
fax +32 2 732 4414
info@gentaur.com
Av. de l' Armée 68
B-1040 Brussels

France

tel 01 43 25 01 50
fax 01 43 25 01 60
9, rue Lagrange
75005 Paris

Italy

tel 02 36 00 65 93
fax 02 36 00 65 94
20135 Milano

Germany

tel +32 16 58 90 45
fax +32 16 50 90 45
Forckenbeckstraße 6
D-52074 Aachen

Japan

tel +81 78 386 0860
fax +81 78 306 0296
Minaatojimaminami-manchi
Chuo-ku, Kobe
065-0047


FIRST STRAND cDNA SYNTHESIS KIT (M-MULV )
Kit Contents:
Components
SD027, 10 reactions
SD028, 20 reactions

10 x Reaction Buffer

50 µl
100 µl

25 mM MgCl2

500 µl
500 µl

dNTP Mix,10mM each

25 µl
50 µl

0.05% [w/v] Gelatin

50 µl
100 µl

Oligo-p(dT)15 Primer, 0.8µg/µl

15 µl
30 µl

Random Primer p(dN)6, 2µg/µl

15 µl
30 µl

RNase Inhibitor, 50 U/µl

15 µl
30 µl

MMLV Reverse Transcriptase

240 U
480 U

RNase-free ddH2O

1.5 ml
1.5 ml

Protocol

1
1
Price
Euro 59
Euro 95
  • It is recommended that the first strand cDNA synthesis is carried out under conditions where RNase contamination has been eliminated. Pipette tips and tubes should be treated with 0.1% diethylpyrocarbonate (DEPC) (soak overnight in 0.1% aqueous solution of DEPC at 37°C, then heat at 10°C for 30 min and autoclave).
  • Wearing gloves is highly recommended.
  • One unit of M-MuLV RT incorporates 1nmole of deoxyribonucleotide into DE81 absorbable form in 10min at 37°C.
  • One unit of ribonuclease inhibitor is defined as the amount of protein required to inhibit 50% of the activity of 5ng of RNase A.

Description:
This kit is designed for preparation of full-length first strand cDNA from RNA templates. The first strand cDNA synthesis kit relies on a cloned enzyme, Moloney murine leukemia virus reverse transcriptase (M-MuLV RT). M-MuLV RT synthesizes first strand cDNA at sites determined by the type of primer used:

  1. random hexamer primer: at non-specific points along an RNA template. In this case, all RNA in a population are templates for cDNA synthesis.
  2. oligo(dT)18: at the 3´-end of poly(A)+ mRNA. In this case, only mRNA with 3´-poly(A) tails are templates for cDNA synthesis.

First strand cDNA synthesized with this system can be used as a template in the polymerase chain reaction (PCR*). As the reaction conditions of cDNA synthesis and PCR* are compatible, cDNA reaction mixture can be added directly to a PCR* mixture. The kit is optimized for maximum yield of full length cDNA. The addition of ribonuclease inhibitor lowers the risk of mRNA degradation during the reaction.
The first strand cDNA synthesized may also be used as a template for second strand synthesis.

*The Polynucleotide Chain reaction (PCR) is covered by the patents owned by Hoffman-La Roche Inc.

References:

  1. Sambrook, J., Fritsch, E.F. and Maniatis, T., Molecular Cloning: A Laboratory Manual (2nd Ed.) Cold Spring Harbor University Press, Cold spring Harbor, NY, 1989.
  2. Ausubel, F.M. et al. eds., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., NY, 1997.
  3. Gerard, G.F., Dlessio, J.M., Meth. in Mol. Biol., V. 16, 73-93, 1993.
 

Home Up

send mail to webmaster@gentaur.com with questions or comments about this web site.
Copyright © 2008 Gentaur Molecular Products
Site powered by Acid Dragon (AC)
Last modified: 05/19/16