Thermo Scientific™ T4 RNA ligasi

Descrizione

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

• The absence of ribonucleases, exodeoxyribonucleases, endodeoxyribonucleases and phosphatases confirmed by appropriate quality tests.

• 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.-[³²P]-(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 MgCl₂, 10mM DTT, 1mM ATP, 10μM 5'-[³²P]-(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 MgCl₂, 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-³²P] 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.