Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase

pfuturbo DNA polymerase for robust, high-fidelity PCR amplification Holly Hogrefe Brad Scott Kirk Nielson Valerie Hedden Connie Hansen Janice Cline Frances Bai Jeff Amberg Ronda Allen Mark Madden Stratagene Stratagene announces the release of pfu...

Transgene in 2007: Partnership and Refinancing Position the Company for its Future Development

...ion per week for 3 weeks of TG4040 at doses of 106 pfu (3 patients), 107 pfu (3 patients) and 108 pfu (9 patients). The patients treated with the higher...

Use of the DCode System to Detect the Food-Borne Bacterial Pathogen Listeria monocytogenes

...ied out in a mixture containing 5 ml of 10x cloned pfu polymerase buffer (Strategene, San Dieg...ir, 1 ml containing 2.5 units of cloned pfu or PfuTurbo (DNA polymerase Strategene, San Diego,...To minimize non-specific annealing of the primers, pfu or PfuTurbo polymerase was added after ...

Optimizing the Direct Amplification of Missing cDNA 5' Ends Using the Eppendorf Mastercycler gradient

...e involved in starch metabolism, was isolated from wheat. A ?ZAPII cDNA library from wheat caryopses was used (approx. 2.5 x 107 pfu per reaction). In addition, the reaction mixture contained primers (0.3 mM each), nucleotides (0.2 mM each), 1.5 mM Mg2+ and Taq...

Optimizing the Direct Amplification of Missing cDNA 5' Ends Using the Eppendorf Mastercycler gradient

...e involved in starch metabolism, was isolated from wheat. A ?ZAPII cDNA library from wheat caryopses was used (approx. 2.5 x 107 pfu per reaction). In addition, the reaction mixture contained primers (0.3 mM each), nucleotides (0.2 mM each), 1.5 mM Mg2+ and Taq...

Lambda DNA Extraction Protocol

... (1% Tryptone, 0.5% yeast extract, 0.5% NaCl, 1 mM MgSO4, pH 7.5) overnight at 37C. Dilute lambda phage stock to l05 pfu per 0.1 ml in SM buffer (0.1 M NaCI, 8 mM MgSO4, 50 mM Tris-Cl, pH 7.5, 0.01% Gelatin), then mix 0.1 ml of the lambda diluti...

Lambda DNA Extraction Protocol

... (1% Tryptone, 0.5% yeast extract, 0.5% NaCl, 1 mM MgSO4, pH 7.5) overnight at 37C. Dilute lambda phage stock to l05 pfu per 0.1 ml in SM buffer (0.1 M NaCI, 8 mM MgSO4, 50 mM Tris-Cl, pH 7.5, 0.01% Gelatin), then mix 0.1 ml of the lambda diluti...

Mammalian Expression Vector for Efficient Cloning of PCR Fragments

...or 25 reactions. Each kit contains predigested pCMV-Script vector, cloned pfu DNA polymerase for polishing, 3 Srf I restriction enzyme, T4 DNA ligase ...ing PCR amplification and to generate blunt-ended DNA fragments, we chose pfu DNA polymerase for PCR amplification of the ampicillin-resistance gen...

Mammalian and Bacterial Expression in One Vector

... Seamless cloning kit 4 was used to amplify the GFP gene from the original TU58 5 construct using high-fidelity pfu DNA polymerase with primers that contained the Eam1104 I recognition site. The digested vector and insert were ligated in the pr...

Unique Enhanced DNA Polymerase Delivers High Fidelity and Great PCR Performance

... proofreading DNA polymerases except Stratagenes pfu and PfuTurbo DNA polymerases. * , Herc...all commercial proofreading DNA polymerases except pfu and PfuTurbo DNA polymerases ( Figure ...mparable or greater accuracy. Only the fidelity of pfu and PfuTurbo DNA polymerases is higher than that...

Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer

... DNA polymerase, a special formulation of cloned pfu DNA polymerase and a novel thermostable PCR-enha...rase amplifies longer targetsin higher yieldthan pfu DNA polymerase (alone), Taq DNA polymerase, and ...ncer and the Turbo additive work in concert with pfu DNA polymerase to provide high product yields wi...

High-Fidelity PCR with a Novel Polymerase Mixture

...ystem consists of a novel mixture of Taq2000 and pfu DNA polymerases and a reaction buffer that has b...tions using long (8 minute) extension times. pfu DNA Polymerase for Highest Fidelity and Thermostability pfu DNA polymerase is purified from the thermophilic a...

Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase

... PfuTurbo DNA Polymerase 1X cloned pfu buffer 1.3 0.2 ... PfuTurbo DNA Polymerase 1.5X cloned pfu buffer 1.1 0.1 ...

Unique DNA Polymerase Formulation Excels in a Broad Range of PCR Applications

...tem. REFERENCES Cline, J., Braman, J.C., and Hogrefe, H.H. (1996) Nucleic Acids Res. 24: 3546-3551. Guide to pfu DNA Polymerase, Stratagene, 1996. Nielson, K.B., et al. (1997) Strategies 10: 29-32. Borns, M., Cline, J., and Hogre...

Generate High-Quality, Directional Plasmid cDNA Libraries

... performed using DNA polymerase I, cloned RNase H 3 and nonmethylated dNTPs. The resulting hemimethylated cDNA is blunted using pfu DNA polymerase, 4 ligated to EcoR I adaptors and digested with Xho I. Since any internal Xho I sites are hemimethylated and pro...

Constructing Directional cDNA Libraries from Limited Amounts of RNA

... numerous advantages over other cloning methods: pfu DNA polymerase, which ensures the highest possible...otides and primers and are treated with pfu DNA polymerase in the presence of dATP....TP, the 3- to 5-exonuclease activity of pfu DNA polymerase removes at least 12 or 1...

Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence

...tion, the PCR product is purified and treated with pfu DNA polymerase in the presence of dATP....CTP, the 3 to 5-exonuclease activity of pfu DNA polymerase removes at least 12 and ...m PCR amplification was gel purified, treated with pfu DNA polymerase at 72C in the presence of dATP an...

Greater Amplification Specificity with New Hot Start PCR Enzyme

...educe nonspecific primer extension by Taq DNA polymerase at low temperatures. One method involves replacing Taq with Stratagenes pfu DNA polymerase, which is less prone to zero-cycle artifacts due to lower activity at nonrestrictive temperatures (e.g., 8% maxim...

High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli

...anded tails, thus allowing for directional cloning. Following PCR amplification , the PCR product is purified, then treated with pfu DNA polymerase in the presence of dATP. The vector and insert DNA are combined, allowed to anneal at room temperature, and trans...

New Protocols for Isolating High- Molecular-Weight Genomic DNA

...action c Average total pfu recoverable d Average ... SCS-8 E. coli strain to determine the number of pfu per reaction. d The estimated total pfu recoverable was determined using the number of...

Performance Comparisons of Commercial RT-PCR Systems

... With the ProSTAR ultra HF RT-PCR system, a versatile two-step format is used, which features PfuTurbo DNA polymerase, 3 a special formulation of pfu DNA polymerase and a novel PCR-enhancing factor*** for amplifying cDNAs with the highest replication fidelity possible. In a previous Strategies a...

pfuturbo DNA Polymerase: A High-Performance, High-Fidelity Enzyme Ideal for PCR Cloning

...A polymerase, 1 a special formulation of cloned pfu DNA polymerase and a novel thermostable factor**...nt-end PCR-Script vector, following treatment with pfu DNA polymerase in the presence of deoxynucleosid...or each enzyme and either PfuTurbo DNA polymerase, pfu DNA polymerase or Taq DNA polymerase and were pe...

prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification

... Frequency (%) pfu 1.3 ... were lowest in amplifications carried out with pfu DNA polymerase ( Table 1 ). ... blend. PfuTurbo DNA polymerase, a blend of cloned pfu DNA polymerase and a novel thermostable PCR enha...

PCR Performance Comparisons Between pfuturbo and Taq DNA Polymerases

...NA polymerase is a special formulation of cloned pfu DNA polymerase and a newly discovered PCR-enhanc...ly higher with PfuTurbo DNA polymerase than with pfu DNA polymerase (alone) or other commercially avail...wo-minute-per-kb extension times recommended for pfu DNA polymerase. 2 In this update, additional ...

Comparing Fidelity and Performance of Proofreading PCR Enzymes

...or high-fidelity PCR applications is Stratagenes pfu DNA polymerase, the most accurate thermostable D... in higher yield than Taq DNA polymerase, cloned pfu DNA polymerase (without enhancer), and other Pfu...er than Pyrococcus furiosus (Pfu), the source of pfu DNA polymerase. KOD DNA polymerase was first des...

Pfu DNA Polymerase as a Hot Start Alternative

...ration prior to the first PCR denaturation step. pfu DNA polymerase, , * which exhibits minimal acti...Taq DNA polymerase-based hot start procedures. pfu DNA polymerase has the lowest error rate of any th... polymerase chain reaction (PCR). Applications for pfu DNA polymerase include gene cloning and expressi...

Enhanced PCR Cloning System

...hough ideal to use with PCR enzymes such as Pfu and pfuturbo DNA polymerases, , ** are versatile and can be adapted for use with any PCR enzyme. pfu and PfuTurbo DNA polymerases are proofreading enzymes that possess 3 - to 5 -exonuclease activity and generate high-fidelity, blunt-end PCR pr...

New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences

... 15 cycles of PCR) and Stratagenes high-fidelity pfu DNA polymerase. After purification of the PCR product, a simple, quick pfu DNA polymerase treatment of the PCR product repl...cation, the PCR product is treated with pfu DNA polymerase in the presence of dATP. In ...

A Positive Selection Assay for Mutation Analysis in Big Blue Animals

...C. Mutant frequency is determined as the number of l cII plaque-forming units (pfu) observed divided by the total pfu screened. The l cII selection scheme has been used to measure mutant frequency inductions in bladder tissue from mice trea...