PDF Datasheetother Pfu DNA Polymerase Recombinant Protein Product Attributes
Product Type: Recombinant Protein
Recombinant Pfu DNA Polymerase based upon sequence from other
Host: QP13023 protein expressed in E. Coli.
Available Tags: Untagged
Protein Construction: A cDNA sequence encoding the sequence of Pfu DNA Polymerase was constructred and used to recombinantly synthesize the protein.
Recommended Applications: See Application Notes
Application Notes
Definition of Unit (for specific activity): 1U of enzyme catalyzes the incorporation of 10nmol of dNTP into acid-insoluble product in 30 minutes at 75°C.
Other 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°C. 7.95% active after 1-hour incubation at 98°C.
Additional Information: 10X Buffer: 200mM Tris-HCl (pH 8.8 at 25°C), 100mM (NH4)2SO4, 100mM KCl, 1% Triton X-100, 1 mg/ml BSA, 20mM MgSO4.
PCR Reaction Protocol: Add the following components to amplify 1kb DNA template: 0.2µl Pfu-DNA Polymerase. 4µl 2.5mM dNTPs. 5µl 10x buffer with MgSO4.1µl Primers mix (10µM each). 1µl Template. 38µl ddH2O. Amplify using the following cycling parameters: Heat Soak: 1 cycle at 94°C/4 min. Denaturation: 30 cycles at 94°C/30 sec. Annealing: 30 cycles at 55°C /30 sec. Extension: 30 cycles at 72°C /90 sec. Final: 1 cycle at 72°C /5 min.
Bioactivity Data: Untested
Amino Acid Sequence: MILDVDYITE EGKPVIRLFK KENGKFKIEH DRTFRPYIYA LLRDDSKIEE VKKITGERHG KIVRIVDVEK VEKKFLGKPI TVWKLYLEHP QDVPTIREKV REHPAVVDIF EYDIPFAKRY LIDKGLIPME GEEELKILAF DIETLYHEGE EFGKGPIIMI SYADENEAKV ITWKNIDLPY VEVVSSEREM IKRFLRIIRE KDPDIIVTYN GDSFDFPYLA KRAEKLGIKL TIGRDGSEPK MQRIGDMTAV EVKGRIHFDL YHVITRTINL PTYTLEAVYE AIFGKPKEKV YADEIAKAWE SGENLERVAK YSMEDAKATY ELGKEFLPME IQLSRLVGQP LWDVSRSSTG NLVEWFLLRK AYERNEVAPN KPSEEEYQRR LRESYTGGFV KEPEKGLWEN IVYLDFRALY PSIIITHNVS PDTLNLEGCK NYDIAPQVGH KFCKDIPGFI PSLLGHLLEE RQKIKTKMKE TQDPIEKILL DYRQKAIKLL ANSFYGYYGY AKARWYCKEC AESVTAWGRK YIELVWKELE EKFGFKVLYI DTDGLYATIP GGESEEIKKK ALEFVKYINS KLPGLLELEY EGFYKRGFFV TKKRYAVIDE EGKVITRGLE IVRRDWSEIA KETQARVLET ILKHGDVEEA VRIVKEVIQK LANYEIPPEK LAIYEQITRP LHEYKAIGPH VAVAKKLAAK GVKIKPGMVI GYIVLRGDGP ISNRAILAEE YDPKKHKYDA EYYIENQVLP AVLRILEGFG YRKEDLRYQK TRQVGLTSWL NIKKS
Reconstitution Instructions: |||
Endotoxin Levels: Not Determined. Testing recommended prior to use in cell culture and can be performed upon request.
Buffer: 50mM Tris-HCl, pH 8.2, 1mM DTT, 0.1mM EDTA, 0.05% CHAPS and 50% glycerol.
Storage Conditions: Pfu DNA Polymerase although stable at 10°C for 5 days, should be stored below -18°C. Please prevent freeze-thaw cycles.
Physical Appearance: Sterile liquid formulation.
| Recombinant other Pfu DNA Polymerase General Information | |
|---|---|
| Alternate Names | |
| DNA polymerase, EC 2.7. 7.7, Pfu polymerase, Pfu-DNA Polymerase. | |
| Curated Database and Bioinformatic Data | |
| UniProt ID(s) | P61875 |
| General Description of Recombinant other Pfu DNA Polymerase. | |
| Pfu DNA Polymerase is a thermo-stable enzyme having a MW of about 90kDa. Pfu DNA Polymerase is derived from E. coli that and cloned from Pyrococcus furiosus strain Vc1 DSM3638. Pfu DNA Polymerase replicates DNA at 75°C, catalyzing the polymerization of nucleotides into duplex DNA in the 5´ to 3´ direction in the existence of magnesium. Pfu DNA Polymerase possesses 3´ to 5´ exonuclease (proofreading) activity. Base misinsertions that take place during polymerization are swiftly removed by the proofreading activity of the polymerase. Therefore, Pfu DNA Polymerase is suggested for use in PCR and primer extension reactions that require high-fidelity synthesis. Pfu DNA Polymerase-generated PCR fragments are blunt-ended. | |
Limitations and Performance Guarantee
enQuire Bio’s products are guaranteed and approved for RESEARCH USE ONLY or manufacturing purposes. This product may not be used as a pharmaceutical agent, or other product intended for human consumption without meeting additional regulatory requirements. This product is guaranteed to work for a period of two years when stored at -70C or colder, and one year when aliquoted and stored at -20C.
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