Hotstart Taq DNA Polymerase

English name：Hotstart Taq DNA Polymerase
Other name: HotStart Taq DNA polymerase

Molecular weight：94kDa，Monomer

Grade：BR

Purity：≥99%
Concentration：5u/ul
Activity definition：The amount of enzyme required for the synthesis of 10 nmol deoxyribonucleic acid into polynucleotide (adsorbed on DE-81) at 74 ℃ for 30 minutes.

Enzyme activity analysis mixture：25mM TAPS(PH9.3,25℃), 50mM KC1, 2mM MgCL2, 0.2mM,various dATPs,dATP,dGTP,dTTP, 0.1mM dCTP, 0.75mM activated salmon essenceDNA和0.4MBq/ML(3H).-dTTP
Save buffer components：20mM Tris-HCL(PH9.0), 1mM DTT, 0.1mM EDTA,100mM KC1, 0.5%(V/V) Tween 20，0.5%(v/v) Nonidet P40和50%(v/v)glycerol

10×Hot start PCR Buffer：200mM Tris-HCL(PH8.3,25℃), 200mM KC1, 50mM (NH4)2SO4
Inhibitor: anionic detergent (when the concentrations of deoxycholic acid, sarcyl and SDS are higher than 0.06%, 0.02 and 0.01%, respectively)

Inactivation: phenol / chloroform extraction

Quality control: Relevant tests show that there is no internal or external deoxyribonuclease and ribonuclease pollution.

Function start:Hot start PCR

Physicochemical properties: Liquid. Designed to enhance the specificity, sensitivity and yield of DNA amplification. The reaction system can be prepared at room temperature with the enzyme, which is convenient to use. It is a chemically modified recombinant Taq DNA polymerase with thermally unstable blocking groups added to the amino acid residues of protein molecules.The enzyme has no activity at room temperature, avoiding the formation of primer dimer and nonspecific extension, so as to increase the specificity of DNA amplification. The activity can be restored by heating at 95 ℃ for 4 minutes.The activated enzyme has the same function as Taq DNA polymerase: it catalyzes the synthesis of DNA in the 5-3 direction, has no detectable 3-5 correction exonuclease activity, and has low 5-3 exonuclease activity.The enzyme also has deoxyribonuclease transfer activity. 11 adenine were added to the 3-terminal of PCR product, and these activities could not be detected before activation.

Purpose: Biochemical research. High yield amplification of complex templates; The specificity of PCR was high - reducing the mismatch effect and the formation of primer dimer; Enhanced PCR sensitivity; It is easy to use and the PCR reaction system is prepared at room temperature; The amplified product has a 3-da protruding end; Hot start PCR; High yield expanded to 3 KB fragment; RT-PCR； Specific amplification of complex cDNA and genomic templates; Amplify low copy DNA template; Real time PCR; Multiplex PCR; PCR products for TA cloning were prepared.