PCR Enzymes

Description

EasyTaq^® DNA Polymerase is purified from E. coli expressing a cloned DNA polymerase from Thermus aquaticus. The enzyme consists of a single polypeptide with a molecular weight of approximately 94 kDa. EasyTaq^® DNA Polymerase has 5′-3′ DNA polymerase activity and 5′-3′ exonuclease activity. It lacks 3′-5′ exonuclease activity. EasyTaq^® DNA Polymerase is suitable for routine amplification. PCR products are unsuitable for PAGE.

Highlights

• Extension rate is about 1-2 kb/min.
• Template-independent “A” can be generated at the 3′ end of the PCR product. PCR products can be directly cloned into pEASY^®-T vectors.
• Amplification of genomic DNA fragment up to 4 kb.

Applications

• Routine PCR
• Colony PCR

Unit Definition

One unit of EasyTaq^® DNA Polymerase incorporates 10 nmol of deoxyribonucleotide into acid-precipitable material in 30 minutes at 74?.

Quality Control

EasyTaq^® DNA Polymerase has passed the following quality control assays: functional absence of double- and single-strand endonuclease activity; >99% homogeneous measured by SDS-PAGE. Each batch of EasyTaq^® DNA Polymerase has been assayed for amplification efficiency to amplify p53 gene from 10 ng of human genomic DNA.

Storage Buffer

20 mM Tris-HCl (pH 8.0), 0.1 mM EDTA, 1 mM DTT, 100 mM KCl, 50% glycerol, stabilizers.

10×EasyTaq^® Buffer (with Mg²⁺)

200 mM Tris-HCl (pH 8.3), 200 mM KCl, 100 mM (NH₄)₂SO₄, 20 mM MgSO₄, others.