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Q5 High Fidelity DNA Polymerase

 

Q5 High-Fidelity DNA Polymerase sets a new standard for both high fidelity and robust performance. The Q5 buffer system is designed to provide superior performance with minimal optimization across a broad range of amplicons regardless of GC content.

"For sure we will use Q5 in the future!"

- Mrs. Evy Vanderheyden

Mrs. Evy Vanderheyden, working at the VIB/KULeuven Molecular Microbiology and Biotechnology department shares her positive experience with Q5 High-Fidelity DNA Polymerase for the robust amplification of genomic yeast DNA:

“Most important with doing PCR for us is a specific PCR product and a high yield. We need a proofreading enzyme since we sequence the obtained PCR products afterwards to check the correct sequences. We do not want errors introduced by the polymerase resulting in false positives or negatives.

For these PCR reactions we used ethanol precipitated genomic yeast DNA. We did a reaction with Phusion High- Fidelity DNA Polymerase like we used to do and compared this with the new Q5 polymerases with the same reaction conditions. It is clear to see that the yield obtained with Q5 High-Fidelity DNA Polymerase (both normal and Hot Start) is much better than the yield obtained with Phusion. After sequencing it turned out to be perfectly PCRed.”

Reaction Set-up

Component 25 µl Reaction Final Concentration

5x Q5 Reaction Buffer

5 µl

1x

10 mM dNTP per nucleotide

0.5 µl each (2 µl total)

800 µM each (200 µM total)

10 μM Forward primer

1.25 µl

0.5 µM

10 µM Reverse primer

1.25 µl

0.5 µM

Template DNA

1 µl

70 ng

Q5 High-Fidelity DNA Polymerase

0.25 µl

0.02 U/µl

Nuclease-Free Water

14.25 µl

 

PCR Protocol

Step Temperature Time

Pre-denaturation

98 °C

30 seconds

30 Cycles *

98 °C

15 seconds

58 °C

30 seconds

72 °C

30 seconds

Final extension 72 °C 120 seconds

Hold

10 °C

Ordering Information

Cat. # Description Quantity

M0491S/L/XL

Q5™ High-Fidelity DNA Polymerase

100/500/2500 units

M0492S/L

Q5™ High-Fidelity 2X Master Mix

100/500 rxns

M0493S/L/XL

Q5™ Hot Start High-Fidelity DNA Polymerase

100/500/2500 units

M0494S/L

Q5™ Hot Start High-Fidelity 2X Master Mix

100/500 rxns

E0555S/L

Q5™ High-Fidelity PCR Kit

50/250 rxns

B9027S

Q5™ Reaction Buffer Pack

6 ml

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