HCY™ Pfu DNA Polymerase Kit

HCY™ Pfu DNA Polymerase Kit

Because Pfu has 3′ to 5′ exonuclease activity, the chance of error during PCR amplification is greatly reduced, and the fidelity is 10 times higher than that of Taq DNA polymerase. The error rate of Pfu is much lower than Taq but also lower than that of some other high fidelity DNA polymerases, such as Vent, DeepVent, Pwo, Tli, etc. Therefore, Pfu is often the first choice of high fidelity DNA polymerases. Therefore, Pfu is often preferred as a cost-effective high-fidelity DNA polymerase.
HCY™ Pfu DNA polymerase has high amplification efficiency and fast amplification speed (4 kb/min, 8 times faster than ordinary Pfu enzyme), which overcomes the defects of ordinary Pfu enzyme’s low amplification efficiency, low yield and slow amplification speed, and greatly shortens the reaction time.

Source

HCY™ Pfu DNA Polymerase is a Recombinant Pfu DNA Polymerase, which is genetically modified by mutation and recombination and purified by expression in E. coli.

Uses

• High-fidelity PCR, point mutation, double-flat-end PCR cloning, and so on.
• The DNA fragments amplified by the Pfu enzyme are double flat-end, which can be used for double flat-end cloning but not for conventional T vector cloning.
• dUTP and dITP or primers containing dUTP and dITP cannot be used in Pfu enzyme-mediated PCR amplification reactions.

Activation Definition

Using activated salmon sperm DNA as template/primer, the activity of 10 nmol of whole nucleotides ingested as acid insoluble was defined as 1 unit of activity (U) at 74°C for 30 min.

Performance of HCY™ Pfu DNA Polymerase

HCY™ pfu DNA Polymeras Amplification Fragment Lengths

• HCY™ pfu DNA Polymeras can amplify 0.5kb/1kb/2kb/5kb fragments of different lengths of λDNA (0-5 kb) with high efficiency.
• M indicates DL10,00 DNA Marker.

HCY™ Pfu DNA Polymeras Amplification Sensitivity

• Amplification of E. coli 300bp fragment with the addition of 0pg/10pg/100pg /1ng/10ng/100ng concentration of template; it can be seen that this product can amplify E. coli genes as low as 1ng template amount.
• NC indicates negative control for HCY™ Pfu DNA Polymerase amplification without template.
• M indicates DL500 DNA Marker.

HCY™ Pfu DNA Polymeras Amplification Speed

• The extension time was 30 sec./60 sec./120 sec. for amplification of 0.5kb/1kb/2kb/5kb size target fragments;
• It can be seen that the extension time of 30 sec. and 120 sec. can well amplify 2kb and 5kb DNA fragments, respectively, indicating that the PCR reaction system with λ DNA as template can set the extension time according to 3 kb/min.
• M indicates DL10,000 DNA Marker.

Quality Control

• SDS-PAGE electrophoretic purity greater than 98%;
• Functional test: sensitivity, specificity, reproducibility of PCR;
• No exogenous nuclease activity, no exogenous endonuclease, and exonuclease contamination;
• No expression of host nucleic acid residue.

Precautions: This product is for scientific research only and cannot be used for medical or diagnostic procedures in humans or animals or as food, cosmetics or household products. Without written permission, authorization, or approval, this product may not be manufactured, licensed for sale, sold, imported or exported, or use all related patents and trademarks of the product.

Packing List

Storage Conditions

For long term storage, please store at -20℃ away from light. If frequent use is required, it can be divided and stored at 4℃ to avoid repeated freezing and thawing. After thawing, it should be mixed well to avoid generating a lot of air bubbles.