Reaction Mixture for Recommended PCR assay:

Component Amount per reaction

DNA template variable

10 M forward Primer 1 L

10 M reverse Primer 1 L

10 mM each dNTP 1 L

5X PerfectRead reaction buffer 10 L

Description:
PerfectRead DNA polymerase 0.5 L
PerfectRead DNA polymerase is a Pfu-based DNA
Nuclease-free water to 50 L
polymerase specially engineered by TEN GIGA BIO.
It was designed to apply in the replication of DNA Total volume 50 L

sequence required high fidelity and high efficiency.

Like Pfu, PerfectRead also exhibits 3'5' Template preparation:
exonuclease activity that provides the proofreading
ability and results in higher fidelity during DNA DNA Amount

synthesis. Moreover, PerfectRead was engineered Genomic DNA 10 ng - 250 ng

to have a better processivity than normal Pfu. Plasmid DNA 1 pg - 10 ng

Therefore, PerfectRead can easily amplify up to 10 cDNA 1 ng - 100 ng
kb blunt-ended DNA fragment from variant
template. All of TEN GIGA BIOs enzymes were Thermocycling Conditions for a Routine PCR:
highly purified by multiple chromatography
procedures that ensure our product free of The recommended parameters may be optimized
contaminant DNA and reduce unexpected results. In for each new primer-template pair for optimal
short, PerfectRead DNA polymerase is an ideal tool specificity and amplification.
for molecular cloning containing long or
complicated amplicons. Segment Number Step Temperature Time

of
Materials supplied: cycles
1. PerfectRead DNA polymerases (2 units/L) 1 1 Initial 95oC 1-5
in storage buffer. denaturation minutes
2. 5X PerfectRead buffer (1 mL/vial)
2 25-35 Denaturation 95oC 10-30
2+
provides a final 2 mM Mg concentration
seconds
in the PCR reaction.
Annealing 45-72oC 10-30

seconds
Applications:
Extension 72oC 15-30
1. High fidelity PCR.
seconds/kb
2. Routine PCR amplification of DNA fragments
3 1 Final 72oC 5-10
up to 10 kb.
Extension minutes
3. Generation of PCR products for cloning and
expression. Hold 14oC --