Certificate of Analysis and Data Sheet

Source:
E. coli

Catalog No.
ENZ-292

Description :

Recombinant Pfu-Plus DNA Polymerase is an enhanced version of Pfu DNA polymerase for high-fidelity PCR. Recombinant Pfu-plus DNA polymerase is a mixture of cloned pyrococcus furiosus DNA polymerase and dUTPase that enhances PCR product yields and increases target length capability without altering DNA replication fidelity. Pfu-plus DNA polymerase can be used to amplify complex genomic DNA targets up to 19kb and vector targets up to 15kb in length.

Concentration:

2.5U/痞.

Purity:

Extensively tested for PCR applications.

Unit Definition:

One unit of enzyme catalyzes the incorporation of 10 nanomoles of deoxyribonucleotides (dNTP) into a polynucleotide fraction (adsorbed on DE-81) in 30min at 72蚓.

Applications:

1. Ideal for high-fidelity amplification. 2. 3'-5' exonuclease activity provides a low error rate. 3. One of the most thermostable DNA polymerases known. 4. Lack of extendase activity means no unwanted 3?overhangs. 5. Optimal for blunt-end PCR cloning. 6. Optimum temperature near 75蚓. 7. 95% active after 1-hour incubation at 98蚓.

Storage Buffer:

20mM Tris-HC1 (pH 8.2), 1mM DTT, 0.1mM EDTA, 100mM KC1, 0.1% Nonidet P40, 0.1% Tween 20 and 50% glycerol.

Stability:

Pfu although stable at 4⁰C for 2 weeks, should be stored desiccated below -18⁰C. Please prevent freeze-thaw cycles.

10X PCR Buffer with MgSO4:

200mM Tris-HC1 ( pH 8.8 at 25蚓), 100mM (NH4)2SO4, 100mM KC1, 1% Triton X-100, 1mg/ml BSA, 20mM MgSO4.

Usage:

This material is offered by ProSpec-TechnoGene for research, laboratory or further evaluation purposes.

Latest Publications:

1. Addendum: Site-directed mutagenesis using Pfu DNA polymerase and T4 DNA ligase.
Biotechniques 2005 Sep;39(3):328
2. Site-directed mutagenesis using Pfu DNA polymerase and T4 DNA ligase.
Biotechniques 2005 Jun;38(6):864, 866, 868
3. A method for filling in the cohesive ends of double-stranded DNA using Pfu DNA polymerase.
Biotechnol Appl Biochem 2005 Dec;42(Pt 3):223-6
4. Improved antibiotic-resistance cassettes through restriction site elimination using Pfu DNA polym erase PCR.
Biotechniques 1998 Nov;25(5):772-4, 776
5. Fidelity and mutational spectrum of Pfu DNA polymerase on a human mitochondrial DNA sequence.
Genome Res 1997 Aug;7(8):843-52
6. PCR fidelity of pfu DNA polymerase and other thermostable DNA polymerases.
Nucleic Acids Res 1996 Sep 15;24(18):3546-51