Thermo Scientific™ T4 RNA Ligase (10 U/μL)

Description

• T4 RNA Ligase catalyzes the ATP-dependent intra- and intermolecular formation of phosphodiester bonds between 5'-phosphate and 3'-hydroxyl termini of oligonucleotides, single-stranded RNA and DNA
• The minimal substrate is a nucleoside 3',5'-biphosphate in intermolecular reaction and oligonucleotide of 8bases in intramolecular reaction
• Source: E
• coli cells with a cloned gene 63 of bacteriophage T4 Molecular Weight: 43.6 kDa monomer Quality Control: The absence of ribonucleases, exodeoxyribonucleases, endodeoxyribonucleases and phosphatases confirmed by appropriate quality tests
• Source: E.coli cells with a cloned gene 63 of bacteriophage T4 Molecular Weight: 43.6kDa monomer Definition of Activity Unit: One unit of the enzyme catalyzes the conversion of 1nmol of 5ft.-[ 32 P]-(A) 12-18 to a phosphatase-resistant form in 30 min
• at 37°C Enzyme activity is assayed in the following mixture: 50mM Tris-HCl (pH 7.5), 10mM MgCl 2 , 10mM DTT, 1mM ATP, 10μM 5'-[ 32 P]-(A) 12-18 (10μM in 5ft.-termini) Storage Buffer: The enzyme is supplied in:20mM Tris-HCl (pH 7.5), 1mM DTT, 50mM KCl, 0.1mM EDTA, 0.03% (v/v) ELUGENT Detergent and 50% (v/v) glycerol 10X Reaction Buffer: 500mM Tris-HCl (pH 7.5 at 25°C), 100mM MgCl 2 , 100mM DTT, 10 mM ATP Inhibition and Inactivation: Inhibitors: metal chelators, SH group-modifying reagents (8)
• Inactivated by heating at 70°C for 10min
• Recommended for: RNA 3'-end labeling with cytidine 3',5'-bis [alpha- 32 P] phosphate (1); Joining RNA to RNA (2); Synthesis of oligoribonucleotides and oligodeoxyribonucleotides (3, 4); Specific modifications of tRNAs (5); Oligodeoxyribonucleotide ligation to single-stranded cDNAs for 5ft
• RACE (Rapid Amplification of cDNA Ends) (6); Site-specific generation of composite primers for PCR (7) Note: The recommended BSA concentration in the reaction mixture is 0.1mg/ml.