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
T4 DNA Ligase (isolated and purified from an E. coli strain) catalyzes the formation of a phosphodiester bond between juxtaposed5’-phosphate and 3’-hydroxyl groups in duplex DNA or RNA. This enzyme is capable of joining adjacent nucleotides in either a blunt-ended or cohesive ended configuration, as well as repairing single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.
cat. no.
amount
note
STS-MSXw10
100ul
M-MLV in different tube, w/o RNase
FOR RESEARCH USE ONLY
STS-MSXw100
4 x 100ul
M-MLV in different tube, w/o 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-MSXw10
STS-MSXw100
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
-
-
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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