Taq-B DNA Polymerase

For routine PCR amplification of DNA fragments up to 5 kb

Products

This product is available worldwide except in China. Kindly note that the delivery takes two to three weeks at the moment. For OEM orders in all countries, including China, please visit our OEM pages.We aim to make this product available globally and speed up delivery. We will keep you posted with any development. Thank you for choosing us as your molecular biology partner.
The Taq-B DNA Polymerase is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention or treatment of a disease, nor has it been validated for such use either alone or in combination with other products.
Image
Taq-B DNA Polymerase (10,000 U)

Cat. No. / ID:  P7250L

10,000 U of Taq-B Polymerase (2 mL at 5000 U/mL) and 10x PCR Buffer. 
Copy order details
NOK 4,480.00

Features

  • Thermally stable, processive 5′→3′ DNA polymerase
  • Possesses an inherent 5′→3′ nick-translation moiety
  • Excels at amplifying shorter (<5 kb) sequences from low-complexity template sources
  • Produces robust yields with little or no optimization of reaction conditions

Product Details

Taq-B DNA Polymerase is a thermally stable, processive, 5′→3′ DNA polymerase from the thermophilic organism Thermus aquaticus YT-1. The 94 kDa protein possesses an inherent 5′→3′ exonuclease activity and lacks a 3′→5′ proofreading function.

Taq excels at amplifying shorter (<5 kb) sequences from low complexity template sources and produces robust yields with little or no optimization of reaction conditions.

The enzyme is supplied in 20mM Tris-HCl, 100mM NaCl, 1mM DTT, 0.1mM EDTA, Stabilizer, 50% glycerol pH 7.5 at 25⁰C.


It is supplied with a 10X PCR Buffer l (B7030) containing: 100mM Tris-HCl, 500mM KCl, 15mM MgCl2 pH 8.3 at 25⁰C.

 

Performance

Assay Specification
Purity >99%
Specific activity 74,625 U/mg
Single-stranded exonuclease 50 U; functional
Double-stranded exonuclease 50 U; functional
Double-stranded endonuclease 50 U; no conversion
E. coli DNA contamination 50 U; <10 copies

 

 

Principle

The enzyme catalyzes 5'→3' synthesis of DNA, has no detectable 3'→5' exonuclease (proofreading) activity and possesses 5'→3' exonuclease activity.

Procedure

Instructions for using Taq-B DNA Polymerase are provided in the corresponding kit protocol in the resources below.


Quality Control

Unit activity was measured using a two-fold serial dilution method. Dilutions of the enzyme were made in 1X reaction buffer and added to 50 μL reactions containing calf thymus DNA, 25 mM TAPS (pH 9.3), 50 mM KCl, 2.0mM MgCl2, 1 mM DTT, 3H-dTTP and 100 μM dNTPs. Reactions were incubated for 10 minutes at 75°C, plunged on ice, and analyzed using the method of Sambrook and Russell (Molecular Cloning, v3, 2001, pp. A8.25-A8.26).


Protein concentration was determined by OD280 absorbance.


Physical purity was evaluated by SDS-PAGE of concentrated and diluted enzyme solutions followed by silver-stain detection. Purity is assessed by comparing the aggregate mass of contaminant bands in the concentrated sample to the mass of the protein of interest band in the diluted sample.


Single-stranded exonuclease was determined in a 50 μL reaction containing a radiolabeled single-stranded DNA substrate and 10 μL of enzyme solution incubated for 4 hours at 37°C.


Double-stranded exonuclease was determined in a 50 μL reaction containing a radiolabeled double-stranded DNA substrate and 10 μL of enzyme solution incubated for 4 hours at 37°C.


Double-stranded endonuclease was determined in a 50 μL reaction containing 0.5 μg of plasmid DNA and 10 μL of enzyme solution incubated for 4 hours at 37°C.


E.coli contamination was evaluated using 5 μL replicate samples of enzyme solution denatured and screened in a TaqMan qPCR assay for the presence of contaminating E.coli genomic DNA using oligonucleotide primers corresponding to the 16S rRNA locus.

 

Applications

This product is available for molecular biology applications such as:

  • Routine PCR amplification of DNA fragments up to 5 kb
  • In vitro amplification
  • Cloning and genotyping

Resources

Protocol Files (1)
Safety Data Sheets (1)