two step amplification phusion flash dna polymerase  (Thermo Fisher)


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    Thermo Fisher two step amplification phusion flash dna polymerase
    Two Step Amplification Phusion Flash Dna Polymerase, supplied by Thermo Fisher, 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/two step amplification phusion flash dna polymerase/product/Thermo Fisher
    Average 88 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    two step amplification phusion flash dna polymerase - by Bioz Stars, 2020-04
    88/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide–RNAs
    Article Snippet: The H1::gRNA scaffold::pol III terminator sequence was then TOPO cloned into pCR4-Blunt (Invitrogen), and sequenced verified; the resulting vector is in the reverse orientation (see below). .. To generate the various gRNAs used in this study, overlapping oligos were annealed and amplified by PCR using two-step amplification Phusion Flash DNA polymerase (Thermo Scientific), and subsequently purified using Carboxylate-Modified Sera-Mag Magnetic Beads (Thermo Scientific) mixed with 2X volume 25%PEG and 1.5M NaCl.

    Amplification:

    Article Title: Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide–RNAs
    Article Snippet: .. To generate the various gRNAs used in this study, overlapping oligos were annealed and amplified by PCR using two-step amplification Phusion Flash DNA polymerase (Thermo Scientific), and subsequently purified using Carboxylate-Modified Sera-Mag Magnetic Beads (Thermo Scientific) mixed with 2X volume 25%PEG and 1.5M NaCl. .. The purified PCR products were then resuspended in H2O and quantitated using a NanoDrop 1000.

    Magnetic Beads:

    Article Title: Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide–RNAs
    Article Snippet: .. To generate the various gRNAs used in this study, overlapping oligos were annealed and amplified by PCR using two-step amplification Phusion Flash DNA polymerase (Thermo Scientific), and subsequently purified using Carboxylate-Modified Sera-Mag Magnetic Beads (Thermo Scientific) mixed with 2X volume 25%PEG and 1.5M NaCl. .. The purified PCR products were then resuspended in H2O and quantitated using a NanoDrop 1000.

    Construct:

    Article Title: Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide–RNAs
    Article Snippet: Plasmid construction To generate the H1 gRNA-expressing construct, overlapping oligos were assembled to create the H1 promoter fused to the 76bp gRNA scaffold and pol III termination signal. .. To generate the various gRNAs used in this study, overlapping oligos were annealed and amplified by PCR using two-step amplification Phusion Flash DNA polymerase (Thermo Scientific), and subsequently purified using Carboxylate-Modified Sera-Mag Magnetic Beads (Thermo Scientific) mixed with 2X volume 25%PEG and 1.5M NaCl.

    Purification:

    Article Title: Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide–RNAs
    Article Snippet: .. To generate the various gRNAs used in this study, overlapping oligos were annealed and amplified by PCR using two-step amplification Phusion Flash DNA polymerase (Thermo Scientific), and subsequently purified using Carboxylate-Modified Sera-Mag Magnetic Beads (Thermo Scientific) mixed with 2X volume 25%PEG and 1.5M NaCl. .. The purified PCR products were then resuspended in H2O and quantitated using a NanoDrop 1000.

    Polymerase Chain Reaction:

    Article Title: Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide–RNAs
    Article Snippet: .. To generate the various gRNAs used in this study, overlapping oligos were annealed and amplified by PCR using two-step amplification Phusion Flash DNA polymerase (Thermo Scientific), and subsequently purified using Carboxylate-Modified Sera-Mag Magnetic Beads (Thermo Scientific) mixed with 2X volume 25%PEG and 1.5M NaCl. .. The purified PCR products were then resuspended in H2O and quantitated using a NanoDrop 1000.

    Generated:

    Article Title: Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide–RNAs
    Article Snippet: To generate the various gRNAs used in this study, overlapping oligos were annealed and amplified by PCR using two-step amplification Phusion Flash DNA polymerase (Thermo Scientific), and subsequently purified using Carboxylate-Modified Sera-Mag Magnetic Beads (Thermo Scientific) mixed with 2X volume 25%PEG and 1.5M NaCl. .. The gRNA-expressing constructs were generated using the Gibson Assembly (NEB) with slight modifications for either the AflII digested plasmid (Addgene #41824) for U6 expression, or BamHI digestion of plasmid just described for H1 expression.

    Expressing:

    Article Title: Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide–RNAs
    Article Snippet: To generate the various gRNAs used in this study, overlapping oligos were annealed and amplified by PCR using two-step amplification Phusion Flash DNA polymerase (Thermo Scientific), and subsequently purified using Carboxylate-Modified Sera-Mag Magnetic Beads (Thermo Scientific) mixed with 2X volume 25%PEG and 1.5M NaCl. .. The gRNA-expressing constructs were generated using the Gibson Assembly (NEB) with slight modifications for either the AflII digested plasmid (Addgene #41824) for U6 expression, or BamHI digestion of plasmid just described for H1 expression.

    Sequencing:

    Article Title: Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide–RNAs
    Article Snippet: The H1::gRNA scaffold::pol III terminator sequence was then TOPO cloned into pCR4-Blunt (Invitrogen), and sequenced verified; the resulting vector is in the reverse orientation (see below). .. To generate the various gRNAs used in this study, overlapping oligos were annealed and amplified by PCR using two-step amplification Phusion Flash DNA polymerase (Thermo Scientific), and subsequently purified using Carboxylate-Modified Sera-Mag Magnetic Beads (Thermo Scientific) mixed with 2X volume 25%PEG and 1.5M NaCl.

    Plasmid Preparation:

    Article Title: Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide–RNAs
    Article Snippet: Paragraph title: Plasmid construction ... To generate the various gRNAs used in this study, overlapping oligos were annealed and amplified by PCR using two-step amplification Phusion Flash DNA polymerase (Thermo Scientific), and subsequently purified using Carboxylate-Modified Sera-Mag Magnetic Beads (Thermo Scientific) mixed with 2X volume 25%PEG and 1.5M NaCl.

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  • Bioz Stars
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  • 88
    Thermo Fisher two step amplification phusion flash dna polymerase
    Two Step Amplification Phusion Flash Dna Polymerase, supplied by Thermo Fisher, 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/two step amplification phusion flash dna polymerase/product/Thermo Fisher
    Average 88 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    two step amplification phusion flash dna polymerase - by Bioz Stars, 2020-04
    88/100 stars
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