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商品編號


ATP



品牌


MCLab



公司


MCLAB.Inc



公司分類(lèi)


DNA/RNA Ligases



Size

10mM

商品信息

T4 RNA Ligase 1 catalyzes the ligation of a 5' phosphoryl-terminated nucleic acid donor to a 3' hydroxyl-terminated nucleic acid acceptor through the formation of a 3'→5' phosphodiester bond, with hydrolysis of ATP to AMP and PP
i
. Substrates include single-stranded RNA and DNA as well as dinucleoside pyrophosphates.




Description:
T4 RNA Ligase 1 catalyzes the ligation of a 5'-phosphoryl-terminated nucleic acid donor to a 3'-hydroxyl-terminated nucleic acid acceptor through the formation of a 3' -5' phosphodiester bond, with hydrolysis of ATP to AMP and PP
i
. Substrates include single-stranded RNA and DNA as well as dinucleoside pyrophosphates.
Application:
- Labeling of 3'-termini of RNA with 5'-[
32
P] pCp
- Inter- and intramolecular joining of RNA and DNA molecules
- Synthesis of single-stranded oligodeoxyribo-nucleotides
- Incorporation of unnatural amino acids into proteins
- Ligation of ssRNA and DNA
Source:
An
E. coli
strain that carries the T4 RNA Ligase 1 gene.
1x T4 RNA Ligase Reaction Buffer (and 10mM ATP):
50 mM Tris-HCl
10 mM MgCl
2
1 mM DTT
pH 7.5 @ 25°C
Heat Inactivation:
65°C for 15 minutes
Heat inactivation by boiling for 2 minutes
Reaction Conditions:
1x T4 RNA Ligase Reaction Buffer
Supplemented with 1 mM ATP
Incubate at 37°C
Unit Definition:
One unit is defined as the amount of enzyme required to convert 1 nmol of 5?-[
32
P]rA16 into a phosphatase-resistant form in 30 minutes at 37°C.
Unit Assay Conditions:
1x T4 RNA Ligase reaction buffer, supplemented with 1 mM ATP, is mixed with the RNA substrate (10?M of 5'-[
32
P]rA16 ) and varying amounts of enzyme. Incubation is at 37°C for 15 minutes.
Concentration:?
10,000 units/ml


Specific Activity:
168,000 U/mg
Storage Conditions:
10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C
Recommended Storage Condition:
-20°C