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
MMLV Mix 5X is is a premixed, ready-to-use reagent (GeneSpin proprietary formulation) containing, dNTPs, MgCl2 , DDT, oligo d(T), random primers RNase Inhibitor and stabilizers optimized for first strand cDNA synthesis. M-MLV Reverse Transcriptase is not supplied directly in the SuperMix 5X but in a different tube.
The MMLV Mix 5X composition is particularly recommended for gene expression analysis in RT-PCR and Real time RT-qPCR.
cat. no.
amount
note
STS-MSX10
100ul
M-MLV in different tube, with RNase
FOR RESEARCH USE ONLY
STS-MSX100
4 x 100ul
M-MLV in different tube, with RNase
SHIPPING
Shipped in green ice.
STORAGE
Store at -20C°
SHELF LIFE
12 months
FORM
Liquid
CONCENTRATION
5X M-MLV Super Mix, 200U/ul M-MLV Reverse Transcriptase (cat no. STS-MRT)
component
STS-MSX10
STS-MSX100
5X M-MLV Buffer
M-MLV Reverse Transcriptase
100ul / 25 reactions
200U/ul / 25ul
4x100ul / 100 reactions
200U/ul / 100ul
RNase free Water
1ml
1ml
RNase
in the 5X M-MLV Buffer tube
in the 5X M-MLV Buffer tube
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:
component
stock conc.
final conc.
20ul reaction
5X M-MLV Buffer
5X
1X
4.0ul
M-MLV Reverse Transcriptase
200U/ul
100-200U
0.5-1ul
RNA
-
-
1ng to 4ug
nuclease free water
-
-
up to 20ul
First Strand cDNA synthesis
Protocol:
Spin down the tubes/plate briefly to remove bubbles.
25°C
5 min
42°C
30 min
85°C
5 min
- Incubate @ 42°C for 60 - 120 min.
- Inactivate the reaction @ 70°C for 10 min.
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