T4 RNA Ligase
Cat. No./Spec.: E1015-A/1,000 U
Concentration: 10 U/µL
Product Description
T4 RNA Ligase catalyzes the ATP-dependent formation of
phosphodiester bonds between 5'-phosphate groups and 3'-hydroxyl
termini of oligonucleotides, single-stranded RNA, and DNA molecules
both intermolecularly and intramolecularly.
Minimum substrates: a nucleotide 3',5'-phosphate in intermolecular
reactions and an 8-nucleotide oligonucleotide in intramolecular
reactions.
Storage Condition & Shelf Life
Store at -20°C.
Source
Recombinant E. coli strain contains the gene 63 cloned from
bacteriophage T4.
Unit Definition
A unit is defined as the amount of enzyme required to catalyze the
conversion of 1 nmol of 5'-[32P]-(A)12-18 to its acid-resistant
form at 37°C within 30 minutes.
Enzyme activity is determined in the following mixture: 50 mM
Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM DTT, 1 mM ATP, and 10 µM
5'-[32P]-(A)12-18 (10 µM at the 5'-terminus).
Scope of Application
- RNA 3'-terminal labeling (using cytidine 3',5'-di[α-32P]
phosphate)
- RNA-to-RNA ligation
- Synthesis of oligoribonucleotides and oligodeoxyribonucleotides
- Specific modification of tRNA
- Ligation of oligodeoxynucleotides to single-stranded cDNA for 5'
RACE (Rapid Amplification of cDNA Ends)
- Site-specific generation of PCR primer complexes
Inhibition and Inactivation
- Inhibitors: metal chelating agents, SH group modifying reagents.
- Heat at 70°C for 10 minutes.
This product is for research use only.
