q5 polymerase master mix  (New England Biolabs)


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    Structured Review

    New England Biolabs q5 polymerase master mix
    Q5 Polymerase Master Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 polymerase master mix/product/New England Biolabs
    Average 95 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    q5 polymerase master mix - by Bioz Stars, 2020-03
    95/100 stars

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    Clone Assay:

    Article Title: Human GPR42 is a transcribed multisite variant that exhibits copy number polymorphism and is functional when heterologously expressed
    Article Snippet: FFAR3 or GPR42 genomic fragments were amplified from 10–15 ng of genomic DNA template in a 25 μL reaction using Q5 polymerase Master mix (NEB). .. Cycling parameters were as follows: 1 hold at 98 °C for 30 s followed by 33 cycles at 98°C for 10 s, 62 °C for 10 s and 72 °C for 90 s, followed by a final extension at 72 °C for 120 s. PCR products were either purified using the Qiafilter PCR Cleanup kit from Qiagen and sequenced (MacrogenUSA) or subcloned using the TOPO PCR blunt cloning kit (Life Technologies) and individual clones were sequenced.

    Amplification:

    Article Title: Changes in the genetic requirements for microbial interactions with increasing community complexity
    Article Snippet: The PCR was performed in a final volume of 20 µL: 10 µL of Q5 polymerase master mix (New England Biolabs), 0.5 µL of forward primer 10 uM, 0.5 µL of reverse primer 10 uM and 5 µL of non-diluted RNA. .. PCR products were run on a 1.7% agarose gel and if genomic DNA was amplified, another DNAse treatment was performed as well as a new verification of absence of genomic DNA.

    Article Title: Human GPR42 is a transcribed multisite variant that exhibits copy number polymorphism and is functional when heterologously expressed
    Article Snippet: .. FFAR3 or GPR42 genomic fragments were amplified from 10–15 ng of genomic DNA template in a 25 μL reaction using Q5 polymerase Master mix (NEB). .. Reactions were assembled in 0.2 mL PCR tubes using an Eppendorf epMotion M5073 liquid handling robot.

    Agarose Gel Electrophoresis:

    Article Title: Changes in the genetic requirements for microbial interactions with increasing community complexity
    Article Snippet: Absence of tRNA and 5S RNA was verified by running 100 ng of RNA on a 1.5% agarose gel and RNA concentration was quantified by nucleic acid quantification in Epoch Microplate Spectrophotometer. .. The PCR was performed in a final volume of 20 µL: 10 µL of Q5 polymerase master mix (New England Biolabs), 0.5 µL of forward primer 10 uM, 0.5 µL of reverse primer 10 uM and 5 µL of non-diluted RNA.

    Polymerase Chain Reaction:

    Article Title: Changes in the genetic requirements for microbial interactions with increasing community complexity
    Article Snippet: .. The PCR was performed in a final volume of 20 µL: 10 µL of Q5 polymerase master mix (New England Biolabs), 0.5 µL of forward primer 10 uM, 0.5 µL of reverse primer 10 uM and 5 µL of non-diluted RNA. .. PCR products were run on a 1.7% agarose gel and if genomic DNA was amplified, another DNAse treatment was performed as well as a new verification of absence of genomic DNA.

    Article Title: Changes in the genetic requirements for microbial interactions with increasing community complexity
    Article Snippet: .. Briefly, PCR was performed in a final volume of 50 µL: 25 µL of Q5 polymerase master mix (New England Biolab), 10 µL of GC enhancer buffer (New England Biolab), 2.5 µL of the common reverse primer (BarSeq_P1 – ) at 10 µM, 2.5 µL of a forward primer from the 96 forward primers (BarSeq_P2_ITXXX) at 10 µM and 50 ng to 2 µg of gDNA. ..

    Article Title: Human GPR42 is a transcribed multisite variant that exhibits copy number polymorphism and is functional when heterologously expressed
    Article Snippet: FFAR3 or GPR42 genomic fragments were amplified from 10–15 ng of genomic DNA template in a 25 μL reaction using Q5 polymerase Master mix (NEB). .. Reactions were assembled in 0.2 mL PCR tubes using an Eppendorf epMotion M5073 liquid handling robot.

    Article Title: Ensemble and Single-Molecule Analysis of Non-Homologous End Joining in Frog Egg Extracts
    Article Snippet: .. 2 × Q5 polymerase master mix (NEB) or similar PCR master mix. .. Taq polymerase should not be used as it will add a nontemplated dA nucleotide to the 3′ end of the PCR product.

    Ethanol Precipitation:

    Article Title: Changes in the genetic requirements for microbial interactions with increasing community complexity
    Article Snippet: The PCR was performed in a final volume of 20 µL: 10 µL of Q5 polymerase master mix (New England Biolabs), 0.5 µL of forward primer 10 uM, 0.5 µL of reverse primer 10 uM and 5 µL of non-diluted RNA. .. According to manufacturer instructions; we used 1 µL of RiboGuard RNAse Inhibitor in each sample as suggested and followed instructions for 1–2.5 ug of RNA input and we used a 2:1 mix of bacterial Ribo-Zero Removal solution and yeast Ribo-Zero Removal solution. rRNA depleted samples were recovered in 10 µL after ethanol precipitation.

    Genomic Sequencing:

    Article Title: Human GPR42 is a transcribed multisite variant that exhibits copy number polymorphism and is functional when heterologously expressed
    Article Snippet: All subsequent primer positions with respect to both FFAR3 and GPR42 genomic sequences are illustrated in . .. FFAR3 or GPR42 genomic fragments were amplified from 10–15 ng of genomic DNA template in a 25 μL reaction using Q5 polymerase Master mix (NEB).

    Spectrophotometry:

    Article Title: Changes in the genetic requirements for microbial interactions with increasing community complexity
    Article Snippet: Absence of tRNA and 5S RNA was verified by running 100 ng of RNA on a 1.5% agarose gel and RNA concentration was quantified by nucleic acid quantification in Epoch Microplate Spectrophotometer. .. The PCR was performed in a final volume of 20 µL: 10 µL of Q5 polymerase master mix (New England Biolabs), 0.5 µL of forward primer 10 uM, 0.5 µL of reverse primer 10 uM and 5 µL of non-diluted RNA.

    Purification:

    Article Title: Changes in the genetic requirements for microbial interactions with increasing community complexity
    Article Snippet: Then, transfer RNAs and 5S RNA were removed using the MEGAclear Kit Purification for Large Scale Transcription Reactions (AMBION, Life Technologies) following manufacturer instructions. .. The PCR was performed in a final volume of 20 µL: 10 µL of Q5 polymerase master mix (New England Biolabs), 0.5 µL of forward primer 10 uM, 0.5 µL of reverse primer 10 uM and 5 µL of non-diluted RNA.

    Article Title: Human GPR42 is a transcribed multisite variant that exhibits copy number polymorphism and is functional when heterologously expressed
    Article Snippet: FFAR3 or GPR42 genomic fragments were amplified from 10–15 ng of genomic DNA template in a 25 μL reaction using Q5 polymerase Master mix (NEB). .. Cycling parameters were as follows: 1 hold at 98 °C for 30 s followed by 33 cycles at 98°C for 10 s, 62 °C for 10 s and 72 °C for 90 s, followed by a final extension at 72 °C for 120 s. PCR products were either purified using the Qiafilter PCR Cleanup kit from Qiagen and sequenced (MacrogenUSA) or subcloned using the TOPO PCR blunt cloning kit (Life Technologies) and individual clones were sequenced.

    Concentration Assay:

    Article Title: Changes in the genetic requirements for microbial interactions with increasing community complexity
    Article Snippet: Absence of tRNA and 5S RNA was verified by running 100 ng of RNA on a 1.5% agarose gel and RNA concentration was quantified by nucleic acid quantification in Epoch Microplate Spectrophotometer. .. The PCR was performed in a final volume of 20 µL: 10 µL of Q5 polymerase master mix (New England Biolabs), 0.5 µL of forward primer 10 uM, 0.5 µL of reverse primer 10 uM and 5 µL of non-diluted RNA.

    Labeling:

    Article Title: Ensemble and Single-Molecule Analysis of Non-Homologous End Joining in Frog Egg Extracts
    Article Snippet: The same fluorescently labeled oligonucleotides used for making the 100bp duplex substrates (see above). .. 2 × Q5 polymerase master mix (NEB) or similar PCR master mix.

    Sequencing:

    Article Title: Changes in the genetic requirements for microbial interactions with increasing community complexity
    Article Snippet: Paragraph title: Library preparation and sequencing ... Briefly, PCR was performed in a final volume of 50 µL: 25 µL of Q5 polymerase master mix (New England Biolab), 10 µL of GC enhancer buffer (New England Biolab), 2.5 µL of the common reverse primer (BarSeq_P1 – ) at 10 µM, 2.5 µL of a forward primer from the 96 forward primers (BarSeq_P2_ITXXX) at 10 µM and 50 ng to 2 µg of gDNA.

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