DNA polymerases were purchased from New England Biolabs (Vent, Vent (exo-), DeepVent (exo-), Bst 2.0, and Therminator) and Thermo Scientific (Taq, Pfu, and Phusion). The reaction mixture (25 μL) contained either 4 μM primer and template or 4 μM hairpin oligonucleotides. The dNTP concentrations were 40 or 200 μM (each) in corresponding buffer. ThermoPol® buffer (New England Biolabs) was used for the Vent, Vent (exo-), DeepVent (exo-), Bst 2.0, and Therminator polymerases. A buffer supplied by the manufacturers was used in the reactions with Taq, Pfu, and Phusion polymerases. One and a half microliters of 25 mM MgCl2were added to the reaction mixtures with Taq polymerase. Two units of enzyme were used for all polymerases, except Bst2.0 (8 units). To initiate pyrophosphorolysis, PPi was added to the reaction mixture at 0.4 mM if not otherwise stated. The reaction mixtures were heated to 95 °C for 1 min and subsequently cooled to 55 °C for 1 min. PE or hairpin modification reactions were performed at 64 or 72 °C for varying amounts of time.
Vent exo
Manufactured by New England Biolabs
The Vent (exo-) is a laboratory equipment designed for controlled venting of gas or liquid from a closed system. It allows for the release of pressure or excess fluid while preventing backflow or contamination. The device operates based on a one-way valve mechanism.
Automatically generated - may contain errors
Lab products found in correlation
Thermopol buffer, by New England Biolabs (1 mentions)
Bst 2, by New England Biolabs (1 mentions)
Therminator, by New England Biolabs (1 mentions)
Deep vent exo, by New England Biolabs (1 mentions)
Bst 3, by New England Biolabs (1 mentions)
Dna oligonucleotide, by Integrated DNA Technologies (1 mentions)
2 protocols using vent exo
1
Comparative Evaluation of DNA Polymerases
2
Comparative DNA Polymerase Characterization
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DNA polymerases: Taq, Vent (exo-), and Bst 3.0 were purchased from New England BioLabs. Taq DNA polymerase is frequently used in methods for amplifying the quantity of short segments of DNA. Vent (exo-) DNA polymerase has been genetically engineered to eliminate the 3´→ 5´ proofreading exonuclease activity associated with Vent DNA Polymerase [20] . This is the preferred form for high yield primer extension reactions. The fidelity of polymerization by this form is reduced to a level of about 2-fold higher than that of Taq DNA Polymerase [21, (link)22] (link). Bst 3.0 DNA polymerase is an in silico designed homolog of Bacillus stearothermophilus DNA polymerase, Large Fragment engineered for improved isothermal amplification performance and increased reverse transcription activity.
DNA oligonucleotides were obtained from Integrated DNA Technologies and Fidelity Systems. All measurements were performed in a buffer solution consisting of 10 mM Tris-HCl at pH 8.7.
DNA oligonucleotides were obtained from Integrated DNA Technologies and Fidelity Systems. All measurements were performed in a buffer solution consisting of 10 mM Tris-HCl at pH 8.7.
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