HiPi Plus DNA Polymerase is suitable for a high fidelity amplification of DNA fragments.This enzyme is designed for a reliable amplification of long (up to 10 kbp for lambda DNA) and complex targets with a robust yield and a high specificity. HiPi Plus DNA Polymerase generates a mixture of PCR products with blunt end and 3´-dA overhangs.
Storage Buffer- unit/㎕ in 20 mM Tris-HCl, pH8.0, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% Tween 20, and 50% Glycerol. Applications- High fidelity PCR for a cloning- RT-PCR- Genomic PCR- Long PCR- Highly specific amplification of DNA fragments with high GC content Unit Definition- One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmole of dNTP into an acid-insoluble form in 30 min at 72℃.- The reaction conditions are : 25 mM TAPS, pH9.3, 50 mM KCl, 2 mM MgCl₂, 1 mM β-mercaptoethanol, 200 μM each dNTPs, 100 μM [α-³²P]dCTP, 12.5 ㎍ activated calf thymus DNA in a total volume of 50 ㎕.10x PCR Reaction Buffer w/ MgCl₂- 500 mM Tris-HCl, pH9.0, 160 mM (NH₄)₂SO₄, 35 mM MgCl₂, 1% Tween 20, 1 mg/ml BSA.10x Q Buffer- PCR additive for increasing specificity of PCR amplification (see figures).
- Q buffer can be added as 0.5x or 1x to PCR mix.