thermopol reaction buffer  (New England Biolabs)


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    Name:
    ThermoPol Reaction Buffer Pack
    Description:
    ThermoPol Reaction Buffer Pack 6 0 ml
    Catalog Number:
    b9004s
    Price:
    22
    Size:
    6 0 ml
    Category:
    Buffers
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    New England Biolabs thermopol reaction buffer
    ThermoPol Reaction Buffer Pack
    ThermoPol Reaction Buffer Pack 6 0 ml
    https://www.bioz.com/result/thermopol reaction buffer/product/New England Biolabs
    Average 99 stars, based on 139 article reviews
    Price from $9.99 to $1999.99
    thermopol reaction buffer - by Bioz Stars, 2020-08
    99/100 stars

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    Polymerase Chain Reaction:

    Article Title: Molecular identification of mosquitoes (Diptera: Culicidae) in southeastern Australia
    Article Snippet: .. The total PCR volume was 25 μ L and consisted of 15.3 μ L 1 × bovine serum albumin (BSA), 2.5 μ L 10 × ThermoPol Reaction Buffer (New England Biolabs, Beverly, MA, USA), 2 μ L 2.5 μ M dNTPs, 1.25 μ L of each 10 μ M/L primer, 0.2 μ L 1.0 U Taq DNA Polymerase, and 2.5 μ L template DNA. ..

    Article Title: PH1: An Archaeovirus of Haloarcula hispanica Related to SH1 and HHIV-2
    Article Snippet: .. To amplify PH1 nucleic acid, approximately 10 ng virus DNA was combined with 500 nM primers, 100 μ M of each dNTP, 1 U Deep Vent DNA Polymerase (New England Biolabs), and 1 × ThermoPol buffer (New England Biolabs), in a total reaction volume of 50 μ L. Template was denatured at 95°C for 10 min, followed by 30 cycles of denaturation at 95°C for 30 sec, annealing at 56°C for 30 sec, extension at 75°C for 2 min, and a final extension at 75°C for 10 min. PCR reactions were performed on a PxE0.2 Thermo Cycler (Thermo Electro Corporation). .. Sequencing reactions using 3.2 pmol primer were performed by the dideoxy chain termination method using ABI PRISM Big Dye Terminator Mix version 3.1 on an ABI 3100 capillary sequencer (Applied Biosystems) by the Applied Genetic Diagnostics Sequencing Service at the Department of Pathology (The University of Melbourne).

    Amplification:

    Article Title: Colorimetric Detection of Ehrlichia Canis via Nucleic Acid Hybridization in Gold Nano-Colloids
    Article Snippet: .. DNA Amplification by LAMP Each 25 μL of LAMP reaction contained a mixture of 1 μL extracted DNA, 40 pmol of each FIP and BIP primer, 5 pmol of each F3 and B3 primer, 5 mM of each dNTP, 3 mM of MgSO4 , ThermoPolTM reaction buffer (20 mM Tris-HCl, 10 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Triton X-100, pH 8.8) Betaine 1.6 M and 8 U of Bst DNA polymerase large fragment (New England Biolabs, Herth, UK). ..

    Size-exclusion Chromatography:

    Article Title: PH1: An Archaeovirus of Haloarcula hispanica Related to SH1 and HHIV-2
    Article Snippet: .. To amplify PH1 nucleic acid, approximately 10 ng virus DNA was combined with 500 nM primers, 100 μ M of each dNTP, 1 U Deep Vent DNA Polymerase (New England Biolabs), and 1 × ThermoPol buffer (New England Biolabs), in a total reaction volume of 50 μ L. Template was denatured at 95°C for 10 min, followed by 30 cycles of denaturation at 95°C for 30 sec, annealing at 56°C for 30 sec, extension at 75°C for 2 min, and a final extension at 75°C for 10 min. PCR reactions were performed on a PxE0.2 Thermo Cycler (Thermo Electro Corporation). .. Sequencing reactions using 3.2 pmol primer were performed by the dideoxy chain termination method using ABI PRISM Big Dye Terminator Mix version 3.1 on an ABI 3100 capillary sequencer (Applied Biosystems) by the Applied Genetic Diagnostics Sequencing Service at the Department of Pathology (The University of Melbourne).

    Plasmid Preparation:

    Article Title: Detection of Bar Transgenic Sugarcane with a Rapid and Visual Loop-Mediated Isothermal Amplification Assay
    Article Snippet: .. The initial LAMP was carried out in a 25.0 μL mixture containing 2.5 μL 10 × ThermoPol Reaction Buffer [New England Biolabs, Massachusetts, USA, including 20.0 mM Tris-HCl (pH 8.8), 10.0 mM KCl, 2.0 mM MgSO4 , 10.0 mM (NH4 )2 SO4 , 0.1% Triton X-100], 3.75 mM MgSO4 (50.0 mM, Sigma-Aldrich Inc., St. Louis, USA), 0.8 μM each FIP and BIP, 0.2 μM each F3 and B3, 0.4 μM each LF and LB, 1.4 mM dNTPs (10.0 mM, TaKaRa Biotechnology Co., Ltd., Dalian, China), 8.0 U Bst DNA polymerase large fragment (New England Biolabs, Massachusetts, USA) and a specified amount of sugarcane genomic DNA or plasmid 1Ac0229. .. The mixture was incubated at 65°C for 60 min, followed by heating at 80°C for 5 min to inactivate the Bst DNA enzyme and terminate the reaction.

    Ligation:

    Article Title: Ancient DNA from Chalcolithic Israel reveals the role of population mixture in cultural transformation
    Article Snippet: .. We filled in the ligated adapters by adding a fill-in reaction mixture (1× ThermoPol buffer (NEB), 250 μM dNTP Mix (ThermoFisher), 0.4 U/μL Bst Polymerase, large fragment (NEB)) to the ligation product, and incubating at 37 °C for 20 min, followed by 80 °C for 20 min. .. Finally, we amplified the libraries via PCR by adding 39 μL of the fill-in reaction product to the PCR reaction mixture (1× Pfu Turbo Cx Reaction Buffer (Agilent Technologies), 0.4 μM PreHyb-F (5′-CTTTCCCTACACGACGCTCTTC-3′), 0.4 μM PreHyb-R (5′-GTGACTGGAGTTCAGACGTGTGCT-3′), 0.2 mM dNTP Mix (ThermoFisher), 5U Pfu Turbo Cx Hotstart DNA Polymerase (Agilent Technologies)).

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  • 90
    New England Biolabs thermophilic dna polymerase buffer
    Ability of mutant Rep proteins to introduce ITR plasmid into AAVS1. Two micrograms of pCMVR78, mutant Rep expression plasmids, or blank vector was transfected into 2 × 10 5 293 cells/well in six-well plates along with 2 μg of pW1, harboring a lacZ expression cassette flanked by ITRs, by a standard calcium phosphate precipitation method. Twenty-four hours later, total cellular <t>DNA</t> was isolated and suspended finally in 200 μl of TE. PCR to detect site-specific integration was carried out as reported previously with minor modifications: 1 μl of isolated genomic DNA was subjected to a thermal cycling reaction in a 20-μl reaction mixture containing <t>1×</t> thermophilic DNA polymerase buffer [10 mM KCl, 20 mM Tris-HCl (pH 8.8), 10 mM (NH 4 ) 2 SO 4 , 4 mM MgSO 4 , 0.1% Triton X-100 (NEB)], 1 μM 5′-CGGCCTCAGTGAGCGAGCGAGC and 5′-CGGGGAGGATCCGCTCAGAGGACA, and 2 U of Deep Vent Exo(−) DNA polymerase (NEB). The cycling conditions were 99°C for 1 min followed by 35 cycles of 99°C for 10 s and 72°C for 4 min. Ten microliters of the PCR mixture was transferred to a hybridization membrane (Hybond-N + ; Amersham) by using a dot blot apparatus and hybridized with a 32 P-labeled AAVS1 probe. The membranes were then analyzed on a BAS-1500 imaging analyzer. The assay was repeated at least four times. p, positive control for hybridization.
    Thermophilic Dna Polymerase Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/thermophilic dna polymerase buffer/product/New England Biolabs
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    thermophilic dna polymerase buffer - by Bioz Stars, 2020-08
    90/100 stars
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    88
    New England Biolabs thermopol pcr buffer
    Ability of mutant Rep proteins to introduce ITR plasmid into AAVS1. Two micrograms of pCMVR78, mutant Rep expression plasmids, or blank vector was transfected into 2 × 10 5 293 cells/well in six-well plates along with 2 μg of pW1, harboring a lacZ expression cassette flanked by ITRs, by a standard calcium phosphate precipitation method. Twenty-four hours later, total cellular <t>DNA</t> was isolated and suspended finally in 200 μl of TE. PCR to detect site-specific integration was carried out as reported previously with minor modifications: 1 μl of isolated genomic DNA was subjected to a thermal cycling reaction in a 20-μl reaction mixture containing <t>1×</t> thermophilic DNA polymerase buffer [10 mM KCl, 20 mM Tris-HCl (pH 8.8), 10 mM (NH 4 ) 2 SO 4 , 4 mM MgSO 4 , 0.1% Triton X-100 (NEB)], 1 μM 5′-CGGCCTCAGTGAGCGAGCGAGC and 5′-CGGGGAGGATCCGCTCAGAGGACA, and 2 U of Deep Vent Exo(−) DNA polymerase (NEB). The cycling conditions were 99°C for 1 min followed by 35 cycles of 99°C for 10 s and 72°C for 4 min. Ten microliters of the PCR mixture was transferred to a hybridization membrane (Hybond-N + ; Amersham) by using a dot blot apparatus and hybridized with a 32 P-labeled AAVS1 probe. The membranes were then analyzed on a BAS-1500 imaging analyzer. The assay was repeated at least four times. p, positive control for hybridization.
    Thermopol Pcr Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/thermopol pcr buffer/product/New England Biolabs
    Average 88 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    thermopol pcr buffer - by Bioz Stars, 2020-08
    88/100 stars
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    89
    New England Biolabs 1x thermopol buffer
    Ability of mutant Rep proteins to introduce ITR plasmid into AAVS1. Two micrograms of pCMVR78, mutant Rep expression plasmids, or blank vector was transfected into 2 × 10 5 293 cells/well in six-well plates along with 2 μg of pW1, harboring a lacZ expression cassette flanked by ITRs, by a standard calcium phosphate precipitation method. Twenty-four hours later, total cellular <t>DNA</t> was isolated and suspended finally in 200 μl of TE. PCR to detect site-specific integration was carried out as reported previously with minor modifications: 1 μl of isolated genomic DNA was subjected to a thermal cycling reaction in a 20-μl reaction mixture containing <t>1×</t> thermophilic DNA polymerase buffer [10 mM KCl, 20 mM Tris-HCl (pH 8.8), 10 mM (NH 4 ) 2 SO 4 , 4 mM MgSO 4 , 0.1% Triton X-100 (NEB)], 1 μM 5′-CGGCCTCAGTGAGCGAGCGAGC and 5′-CGGGGAGGATCCGCTCAGAGGACA, and 2 U of Deep Vent Exo(−) DNA polymerase (NEB). The cycling conditions were 99°C for 1 min followed by 35 cycles of 99°C for 10 s and 72°C for 4 min. Ten microliters of the PCR mixture was transferred to a hybridization membrane (Hybond-N + ; Amersham) by using a dot blot apparatus and hybridized with a 32 P-labeled AAVS1 probe. The membranes were then analyzed on a BAS-1500 imaging analyzer. The assay was repeated at least four times. p, positive control for hybridization.
    1x Thermopol Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 89/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1x thermopol buffer/product/New England Biolabs
    Average 89 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    1x thermopol buffer - by Bioz Stars, 2020-08
    89/100 stars
      Buy from Supplier

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    Ability of mutant Rep proteins to introduce ITR plasmid into AAVS1. Two micrograms of pCMVR78, mutant Rep expression plasmids, or blank vector was transfected into 2 × 10 5 293 cells/well in six-well plates along with 2 μg of pW1, harboring a lacZ expression cassette flanked by ITRs, by a standard calcium phosphate precipitation method. Twenty-four hours later, total cellular DNA was isolated and suspended finally in 200 μl of TE. PCR to detect site-specific integration was carried out as reported previously with minor modifications: 1 μl of isolated genomic DNA was subjected to a thermal cycling reaction in a 20-μl reaction mixture containing 1× thermophilic DNA polymerase buffer [10 mM KCl, 20 mM Tris-HCl (pH 8.8), 10 mM (NH 4 ) 2 SO 4 , 4 mM MgSO 4 , 0.1% Triton X-100 (NEB)], 1 μM 5′-CGGCCTCAGTGAGCGAGCGAGC and 5′-CGGGGAGGATCCGCTCAGAGGACA, and 2 U of Deep Vent Exo(−) DNA polymerase (NEB). The cycling conditions were 99°C for 1 min followed by 35 cycles of 99°C for 10 s and 72°C for 4 min. Ten microliters of the PCR mixture was transferred to a hybridization membrane (Hybond-N + ; Amersham) by using a dot blot apparatus and hybridized with a 32 P-labeled AAVS1 probe. The membranes were then analyzed on a BAS-1500 imaging analyzer. The assay was repeated at least four times. p, positive control for hybridization.

    Journal: Journal of Virology

    Article Title: Charged-to-Alanine Scanning Mutagenesis of the N-Terminal Half of Adeno-Associated Virus Type 2 Rep78 Protein

    doi:

    Figure Lengend Snippet: Ability of mutant Rep proteins to introduce ITR plasmid into AAVS1. Two micrograms of pCMVR78, mutant Rep expression plasmids, or blank vector was transfected into 2 × 10 5 293 cells/well in six-well plates along with 2 μg of pW1, harboring a lacZ expression cassette flanked by ITRs, by a standard calcium phosphate precipitation method. Twenty-four hours later, total cellular DNA was isolated and suspended finally in 200 μl of TE. PCR to detect site-specific integration was carried out as reported previously with minor modifications: 1 μl of isolated genomic DNA was subjected to a thermal cycling reaction in a 20-μl reaction mixture containing 1× thermophilic DNA polymerase buffer [10 mM KCl, 20 mM Tris-HCl (pH 8.8), 10 mM (NH 4 ) 2 SO 4 , 4 mM MgSO 4 , 0.1% Triton X-100 (NEB)], 1 μM 5′-CGGCCTCAGTGAGCGAGCGAGC and 5′-CGGGGAGGATCCGCTCAGAGGACA, and 2 U of Deep Vent Exo(−) DNA polymerase (NEB). The cycling conditions were 99°C for 1 min followed by 35 cycles of 99°C for 10 s and 72°C for 4 min. Ten microliters of the PCR mixture was transferred to a hybridization membrane (Hybond-N + ; Amersham) by using a dot blot apparatus and hybridized with a 32 P-labeled AAVS1 probe. The membranes were then analyzed on a BAS-1500 imaging analyzer. The assay was repeated at least four times. p, positive control for hybridization.

    Article Snippet: One microliter of isolated genomic DNA was subjected to a thermal cycling reaction in a 20-μl reaction mixture containing 1× thermophilic DNA polymerase buffer [10 mM KCl, 20 mM Tris-HCl (pH 8.8), 10 mM (NH4 )2 SO4 , 4 mM MgSO4 , 0.1% Triton X-100 (New England Biolabs {NEB})], 1 μM 5′-CGGCCTCAGTGAGCGAGCGAGC and 5′-CGGGGAGGATCCGCTCAGAGGACA, and 2 U of Deep Vent Exo(−) DNA polymerase (NEB).

    Techniques: Mutagenesis, Introduce, Plasmid Preparation, Expressing, Transfection, Isolation, Polymerase Chain Reaction, Hybridization, Dot Blot, Labeling, Imaging, Positive Control

    EMSA of mutant Rep78 synthesized in vitro. Three microliters of in vitro-synthesized Rep78 or mutant Rep protein in the absence of radiolabeled amino acids was incubated for 15 min at 30°C with 20,000 cpm of 5′-end-labeled RBS oligonucleotide probe in a 10-μl solution of 10 mM HEPES-KOH (pH 7.9), 50 mM KCl, 0.1 mM EDTA, 0.05% BSA, 10% glycerol, and 1 μg of sheared calf thymus DNA. Reaction products were separated on 4% nondenaturing polyacrylamide gels, dried, and then analyzed on a BAS-1500 imaging analyzer. Unprogrammed lysate (no DNA) and lysate programmed with vector (vector) were also included as negative controls. ∗, complete loss of binding activity; ∗∗, consistently lower binding activity compared to the wild type.

    Journal: Journal of Virology

    Article Title: Charged-to-Alanine Scanning Mutagenesis of the N-Terminal Half of Adeno-Associated Virus Type 2 Rep78 Protein

    doi:

    Figure Lengend Snippet: EMSA of mutant Rep78 synthesized in vitro. Three microliters of in vitro-synthesized Rep78 or mutant Rep protein in the absence of radiolabeled amino acids was incubated for 15 min at 30°C with 20,000 cpm of 5′-end-labeled RBS oligonucleotide probe in a 10-μl solution of 10 mM HEPES-KOH (pH 7.9), 50 mM KCl, 0.1 mM EDTA, 0.05% BSA, 10% glycerol, and 1 μg of sheared calf thymus DNA. Reaction products were separated on 4% nondenaturing polyacrylamide gels, dried, and then analyzed on a BAS-1500 imaging analyzer. Unprogrammed lysate (no DNA) and lysate programmed with vector (vector) were also included as negative controls. ∗, complete loss of binding activity; ∗∗, consistently lower binding activity compared to the wild type.

    Article Snippet: One microliter of isolated genomic DNA was subjected to a thermal cycling reaction in a 20-μl reaction mixture containing 1× thermophilic DNA polymerase buffer [10 mM KCl, 20 mM Tris-HCl (pH 8.8), 10 mM (NH4 )2 SO4 , 4 mM MgSO4 , 0.1% Triton X-100 (New England Biolabs {NEB})], 1 μM 5′-CGGCCTCAGTGAGCGAGCGAGC and 5′-CGGGGAGGATCCGCTCAGAGGACA, and 2 U of Deep Vent Exo(−) DNA polymerase (NEB).

    Techniques: Mutagenesis, Synthesized, In Vitro, Incubation, Labeling, Imaging, Plasmid Preparation, Binding Assay, Activity Assay