T4 RNA ligase

T4 RNA ligase

Product description:

English name: T4 RNA Ligase
Molecular Weight: 43.6kDa, monomer

Level: BR
Source: T4 phage clone containing the E. coli gene 63
Concentration: 10u / ul
Activity definition: refers to 37 ℃, over 30 minutes 1nmol 5 '- [32P] - (A) 12-18 into the desired amount of enzyme form phosphatase-resistant
Activity assay mixture: 50mM Tris-HCL (PH7.5), 10mM DTT, 10mM MgCL2, 1mM ATP, 10uM 5 '- [32P] - (A) 12-18 (10uM 5'- terminus)
Preservation solution components: 20mM Tris-HCL (PH7.5), 50mM KC1, 1mM DTT, 0.1mM EDTA, 0.5mM ELUGENT denaturant and 50% (v / v) glycerol
10 * reaction buffer: 500mM HEPES-NaOH (PH8.025 ℃), 100mM MgCL2, 100mM DTT
Inhibitors: metal chelators, SH group modifying agent
Inactivation: 70 ℃ heated for 10 minutes
Quality control: tests show no endo- and exo-DNase, RNase, phosphatase pollution
Note: the concentration of ATP in the reaction system depends on the type of ligation reaction, recommended reaction concentration of BSA is 0.1mg / ml
Properties: between recombinase catalyze ATP-dependent molecules and molecular phosphodiester linkages (5'-phosphate groups and 3'-hydroxyl terminus), the substrate molecule is a polynucleotide, oligonucleotide, ssRNA and ssDNA, the smallest connection between a substrate molecule is a nucleoside 3 ', 5'-diphosphate, and the minimum inner substrate molecule is 8 base oligonucleotides. Supplied with reaction buffer, ATP and the BSA solution. Catalytic 3 '→ 5' phosphodiester bond is formed, so that the 5'-hydroxyl and 5'-phosphate ends of the connection terminal nucleotide. Role substrate comprises single-stranded RNA, DNA and dinucleoside pyrophosphate . Supplied with 10X Reaction Buffer
Use: Biochemical studies connecting RNA and RNA; RNA3'- end-labeled cytosine 3 ', 5'-bis [a-32P] phosphate; specifically modified tRNA; the synthetic oligonucleotides cyclization; oligo deoxyribose. nucleotide single-stranded cDNA connection for 5'RACE (rapid amplification of cDNA ends) Analysis
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