T7 DNA Polymerase

English name：T7 DNA Polymerase

Molecular weight：It consists of two subunits, 804kda polypeptide (T7 phage gene 5 expression product) and 12kda thioredoxin (E. coli trxA gene expression product)

Grade：BR
Source: Two E. coli strains, one containing T7 phage clone gene 5 and the other containing E. coli clone gene trxA

Concentration：10u/ul
Activity definition: The amount of enzyme required to enable 10 nmol of deoxyribonucleotide to be incorporated into polynucleotide fragments (adsorbed on DE-81) within 30 minutes at 37 ° C.

Enzyme activity analysis mixture：40mM Tris-HCL(PH7.5), 1mM DTT, 10mM MgCL2, 0.1mg/ml BSA, 0.033mM:VariousdNTP,0.4MBq/ML(3H)-dTTP和0.5mM:Alkaline denatured bovine thymus DNA

Preservation buffer components: 20 mM potassium phosphate (pH7.4), 0.1 mm EDTA, 1 mm DTT and 50% (V / V) glycerol

10×Reaction buffer：400mM Tris-HCL(PH7.5，25℃), 100mM MgCL2, 10mM DTT
Inhibitors: metal chelating agents, modifying reagents (anhydrous acetic acid and N-ethyl maleimide) can inactivate 3 '→ 5' exonuclease, but do not affect polymerase activity.

Inactivation: Heating at 75 ℃ for 10 minutes

Quality control: The test shows that there is no endodeoxyribonuclease pollution

Note for use: Short incubation is required for analysis at 37 ℃.

Physicochemical properties: Suspension, recombinant enzyme.T7 DNA polymerase is a template dependent DNA polymerase that catalyzes DNA synthesis in the 5 '→ 3' direction.The DNA polymerase has high continuous synthesis ability and can continuously synthesize long fragments of DNA. The enzyme has 3 '→ 5' exonuclease activity for single stranded and double stranded DNA.

Purpose: Biochemical research. Remove the residual genomic DNA and purify the covalently closed-loop DNA molecule; Long fragment primer extension reaction; DNA 3 '- terminal labeling; Chain extension reaction at site directed mutation sites; DNA 5 '- protrusion site flattening; Second strand cDNA synthesis; Apoptosis related DNA fragments were detected in situ