MMLV Mix 5X (with or without RNase Inhibitor) is a premixed, ready-to-use reagent (GeneSpin proprietary formulation) containing, dNTPs, MgCl2 , DDT, oligo d(T), random primers 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
cat. no.
amount
note
STS-MSX 10
100 ul
M-MLV in different tube with RNase
STS-MSXw 10
100 ul
M-MLV in different tube w/o RNase
STS-MSX 100
4x100 ul
M-MLV in different tube with RNase
STS-MSXw 100
4x100 ul
M-MLV in different tube w/o RNase
FOR RESEARCH USE ONLY
SHIPPING
Shipped in green ice.
CONCENTRATION
5X M-MLV Super Mix, 200U/ul M-MLV Reverse Transcriptase (cat no. STS-MRT)
FORM
Liquid
SHELF LIFE
12 months
STORAGE
Store at -20°C
Components
component
STS-MSX 10
STS-MSX 100
STS-MSXw 10
STS-MSXw 100
5X M-MLV Buffer
100 ul/ 25 reaction
4x100 ul/ 100 reactions
100 ul/ 25 reaction
4x100 ul/ 100 reactions
M-MLV reverse transcriptase
RNase free water
RNase
200U/ul - 25ul
1 ml
In the 5X M-MLV buffer
200U/ul - 100ul
1ml
In the 5X M-MLV buffer
200U/ul - 25ul
1 ml
-
200U/ul - 100ul
1ml
-
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
M-MLV reverse transcriptase
RNA
5X
1ug/ul each
-
1X
50ng/ul each
-
4.0ul
0.5-1 ul
1ng to 4ng
RNase free water
-
-
up to20ul
First cDNA synthesis
Spin down the tubes/plate briefly to remove bubbles.
25°C
5min
42°C
30 min
85°C
2 min
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