آنزیم پلیمراز فیوژن | Phusion DNA Polymerase Enzyme

Phusion DNA polymerase

Cat. No:

FPLF004.0250

FPLF004.0500

Contents:

Description:

Phusion DNA Polymerase is a chimeric Pfu which has a DNA binding protein at the N-terminal portion of the gene. This enzyme keeps significant activity after exposure to 99 °C or repeated exposure to 98 °C with more processivity and extention rate than Pfu DNA polymerase. It catalyzes the polymerization of nucleotides into duplex DNA in the 5’_3′ direction, resulting in blunt-ended PCR products without 3′-dA overhangs. Phusion DNA Polymerase exhibits 3’_5′ exonuclease (proofreading) activity that enables the polymerase to correct the mis-incorporation of nucleotide, and lacks 5’_3′ exonuclease activity. It is suitable for PCR and primer extension reaction that requires high fidelity when the PCR fragment is relatively higher than 3 kb. The enzyme exhibits 3´>5´ proofreading activity, resulting in over 20-fold higher PCR fidelity than possible with Taq DNA Polymerases.

Kit storage:

This kit should be stored at -20 ^oC. Under this condition reagents are stable for two years from the date of production.

Protocol:

• Thaw 5X reaction buffer, dNTP mixture.
• Mix the master mix thoroughly and dispense appropriate volumes into PCR tubes or plates.
• Add templates DNA to the individual PCR tubes or wells containing the master mix.
• Program the PCR machine according to the program outlined.

Note:

Longer extension time makes nonspecific bands

Extension rate for this enzyme is near 3000 bp/min.

Agarose gel Electrophoresis:

Run the total 5-7 μL of PCR products alongside 3μL DNA marker on a 2% agarose gel containing Green viewer DNA safe stain.