1x q5 buffer  (New England Biolabs)


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    New England Biolabs 1x q5 buffer
    1x Q5 Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1x q5 buffer/product/New England Biolabs
    Average 93 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    1x q5 buffer - by Bioz Stars, 2020-04
    93/100 stars

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    DNA Extraction:

    Article Title: CRISPR-Mediated Base Editing Enables Efficient Disruption of Eukaryotic Genes through Induction of STOP Codons
    Article Snippet: The cell pellet was resuspended in the Quick Extract DNA Extraction Solution (Epicenter) and heated sequentially at 65°C for 5 min and 95°C for 5 min to isolate genomic DNA (gDNA). .. The following genomic loci were targeted by sgSTOPs for SPRTN (chr1+ 231338561), SMARCAL1 (chr2+ 216414804), CHEK2 (chr22- 28734431), TIMELESS (chr12- 56431493) and FANCM ) using the default parameters with the following changes: Mispriming library = “HUMAN,” Primer size “min = 25, Opt = 27, Max = 30,” Primer Tm “Min= 57.0, Opt = 60.0, Max = 63.0.” PCR reactions were prepared in a 25 μL reaction volume containing: 1 μM forward and reverse primers, 0.1 nM dNTPs (NEB #N0447L), 1X Q5 buffer (NEB), 200–300 ng gDNA, 1 unit Q5 polymerase (NEB) and water.

    Clone Assay:

    Article Title: The Pokeweed Leaf mRNA Transcriptome and Its Regulation by Jasmonic Acid
    Article Snippet: Paragraph title: Cloning of Novel PAP Isoform and PAP Natural Antisense Transcript ... Following cDNA synthesis, a PCR reaction was conducted, containing 1X Q5 buffer (NEB), 0.5 μM forward primer, 0.5 μM reverse primer, 200 mM dNTPs, 2 μL cDNA and 1 unit Q5 DNA polymerase (NEB) in a total volume of 50 μL.

    Transfection:

    Article Title: CRISPR-Mediated Base Editing Enables Efficient Disruption of Eukaryotic Genes through Induction of STOP Codons
    Article Snippet: The occurrence of iSTOP-mediated editing was monitored three days after transfection of HEK293T cells with BE3 and sgSTOPs, as indicated above. .. The following genomic loci were targeted by sgSTOPs for SPRTN (chr1+ 231338561), SMARCAL1 (chr2+ 216414804), CHEK2 (chr22- 28734431), TIMELESS (chr12- 56431493) and FANCM ) using the default parameters with the following changes: Mispriming library = “HUMAN,” Primer size “min = 25, Opt = 27, Max = 30,” Primer Tm “Min= 57.0, Opt = 60.0, Max = 63.0.” PCR reactions were prepared in a 25 μL reaction volume containing: 1 μM forward and reverse primers, 0.1 nM dNTPs (NEB #N0447L), 1X Q5 buffer (NEB), 200–300 ng gDNA, 1 unit Q5 polymerase (NEB) and water.

    Amplification:

    Article Title: Using a soil bacterial species balance index to estimate potato crop productivity
    Article Snippet: Prokaryota (archaea and bacteria) rRNA 16S (V4 region) gene was amplified using 515FB and 806RB primers [ – ] and a two-step dual-indexed PCR approach specifically designed for Illumina instruments by Plateforme d’analyses génomiques (IBIS, Université Laval, Quebec City, QC, Canada). .. Briefly, the gene specific sequence was fused to the Illumina TruSeq sequencing primers and PCR was carried out in a total volume of 50 μl that contained 1X Q5 buffer (NEB, Whitby, ON, Canada), 0.25 μM of each primer, 200 μM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase (NEB, Whitby, ON, Canada), and 1 μl of template DNA.

    Article Title: The hindgut microbiota of praying mantids is highly variable and includes both prey-associated and host-specific microbes
    Article Snippet: During the initial reaction the V4 region of the 16S rRNA gene was amplified with the 515F ( GTGCCAGCMGCCGCGGTAA ) and 806R ( GGACTACHVGGGTWTCTAAT ) primers. .. The initial reaction contained 1X Q5 buffer (New England Biolabs [NEB], Ipswitch, MA), 200 μM dNTPS, 0.5 μM 515F, 0.5 μM 806R, 2 ng template DNA, and 0.02 U/μL Q5 Hot Start High-Fidelity DNA polymerase (NEB) for a total reaction volume of 10 μL.

    Article Title: Direct-fed microbial supplementation influences the bacteria community composition of the gastrointestinal tract of pre- and post-weaned calves
    Article Snippet: Paragraph title: Amplification of bacterial ribosomal DNA and sequencing ... The PCR was carried out in a total volume of 50 µL that contains 1X Q5 buffer (NEB), 0.25 µM of each primer, 200 µM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase and 1 µL of template cDNA.

    Article Title: The hindgut microbiota of praying mantids is highly variable and includes both prey-associated and host-specific microbes
    Article Snippet: During the initial reaction the V4 region of the 16S rRNA gene was amplified with the 515F ( GTGCCAGCMGCCGCGGTAA ) and 806R ( GGACTACHVGGGTWTCTAAT ) primers. .. The initial reaction product was added to the second reaction, which contained 1X Q5 buffer (NEB), 200 μM dNTPS, 0.5 μM 515F, 0.5 μM 806R, and 0.02 U/μL Hot Start High-Fidelity DNA polymerase (NEB) at a volume of 21 μL.

    Agarose Gel Electrophoresis:

    Article Title: Using a soil bacterial species balance index to estimate potato crop productivity
    Article Snippet: Genomic DNA quality was also verified by electrophoresis on a 1.6% (w/w) agarose gel and visualized under UV with a Gel Doc XR+ instrument (Biorad, Hercules, CA., USA). .. Briefly, the gene specific sequence was fused to the Illumina TruSeq sequencing primers and PCR was carried out in a total volume of 50 μl that contained 1X Q5 buffer (NEB, Whitby, ON, Canada), 0.25 μM of each primer, 200 μM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase (NEB, Whitby, ON, Canada), and 1 μl of template DNA.

    Article Title: The hindgut microbiota of praying mantids is highly variable and includes both prey-associated and host-specific microbes
    Article Snippet: The initial reaction product was added to the second reaction, which contained 1X Q5 buffer (NEB), 200 μM dNTPS, 0.5 μM 515F, 0.5 μM 806R, and 0.02 U/μL Hot Start High-Fidelity DNA polymerase (NEB) at a volume of 21 μL. .. PCR conditions were 98°C for 30 s for initial denaturation; 4 cycles at 98°C for 10 s, 52°C for 10 s, and 72°C for 30 s; 6 cycles at 98°C for 10 s and 72°C for 1 min; and the final extension step at 72°C for 2 min. Amplicons were visualized on a 2% w/v agarose gel.

    Isolation:

    Article Title: CRISPR-Mediated Base Editing Enables Efficient Disruption of Eukaryotic Genes through Induction of STOP Codons
    Article Snippet: The isolated gDNA was quantified using Nanodrop, diluted in water and stored at −20°C or directly used in PCR reactions. .. The following genomic loci were targeted by sgSTOPs for SPRTN (chr1+ 231338561), SMARCAL1 (chr2+ 216414804), CHEK2 (chr22- 28734431), TIMELESS (chr12- 56431493) and FANCM ) using the default parameters with the following changes: Mispriming library = “HUMAN,” Primer size “min = 25, Opt = 27, Max = 30,” Primer Tm “Min= 57.0, Opt = 60.0, Max = 63.0.” PCR reactions were prepared in a 25 μL reaction volume containing: 1 μM forward and reverse primers, 0.1 nM dNTPs (NEB #N0447L), 1X Q5 buffer (NEB), 200–300 ng gDNA, 1 unit Q5 polymerase (NEB) and water.

    Spectrophotometry:

    Article Title: Using a soil bacterial species balance index to estimate potato crop productivity
    Article Snippet: The quality and quantity of the DNA extracts were evaluated by spectrophotometry using a Biophotometer (Eppendorf, Mississauga, ON, Canada) with a G1.0 microcuvette μCuvette (Eppendorf, Mississauga, ON, Canada) with readings at 260, 280, 230, and 320 nm. .. Briefly, the gene specific sequence was fused to the Illumina TruSeq sequencing primers and PCR was carried out in a total volume of 50 μl that contained 1X Q5 buffer (NEB, Whitby, ON, Canada), 0.25 μM of each primer, 200 μM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase (NEB, Whitby, ON, Canada), and 1 μl of template DNA.

    Article Title: Direct-fed microbial supplementation influences the bacteria community composition of the gastrointestinal tract of pre- and post-weaned calves
    Article Snippet: The PCR was carried out in a total volume of 50 µL that contains 1X Q5 buffer (NEB), 0.25 µM of each primer, 200 µM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase and 1 µL of template cDNA. .. Quality of the purified PCR product was checked on a DNA7500 BioAnalyzer chip (Agilent) and quantified using a Nanodrop 1000 Spectrophotometer (Thermo Fisher Scientific).

    Article Title: Exacerbation induces a microbiota shift in sputa of COPD patients
    Article Snippet: PCR reactions contained: 1X Q5 buffer (New England Biolabs, Ipswich, Massachusetts, USA), 200 μM of each dNTP (Feldan, Quebec, Quebec, Canada), 0.2 μM of each 454 primer (IDT), 1 U of Q5 High-Fidelity DNA polymerase (NEB), and 1 μL of template DNA (10–50 ng). .. PCR products were then quantified with a NanoDrop 2000 spectrophotometer (Thermo Scientific).

    RFLP Assay:

    Article Title: CRISPR-Mediated Base Editing Enables Efficient Disruption of Eukaryotic Genes through Induction of STOP Codons
    Article Snippet: Paragraph title: RFLP assay and DNA sequencing ... The following genomic loci were targeted by sgSTOPs for SPRTN (chr1+ 231338561), SMARCAL1 (chr2+ 216414804), CHEK2 (chr22- 28734431), TIMELESS (chr12- 56431493) and FANCM ) using the default parameters with the following changes: Mispriming library = “HUMAN,” Primer size “min = 25, Opt = 27, Max = 30,” Primer Tm “Min= 57.0, Opt = 60.0, Max = 63.0.” PCR reactions were prepared in a 25 μL reaction volume containing: 1 μM forward and reverse primers, 0.1 nM dNTPs (NEB #N0447L), 1X Q5 buffer (NEB), 200–300 ng gDNA, 1 unit Q5 polymerase (NEB) and water.

    Electrophoresis:

    Article Title: Using a soil bacterial species balance index to estimate potato crop productivity
    Article Snippet: Genomic DNA quality was also verified by electrophoresis on a 1.6% (w/w) agarose gel and visualized under UV with a Gel Doc XR+ instrument (Biorad, Hercules, CA., USA). .. Briefly, the gene specific sequence was fused to the Illumina TruSeq sequencing primers and PCR was carried out in a total volume of 50 μl that contained 1X Q5 buffer (NEB, Whitby, ON, Canada), 0.25 μM of each primer, 200 μM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase (NEB, Whitby, ON, Canada), and 1 μl of template DNA.

    Polymerase Chain Reaction:

    Article Title: CRISPR-Mediated Base Editing Enables Efficient Disruption of Eukaryotic Genes through Induction of STOP Codons
    Article Snippet: .. The following genomic loci were targeted by sgSTOPs for SPRTN (chr1+ 231338561), SMARCAL1 (chr2+ 216414804), CHEK2 (chr22- 28734431), TIMELESS (chr12- 56431493) and FANCM ) using the default parameters with the following changes: Mispriming library = “HUMAN,” Primer size “min = 25, Opt = 27, Max = 30,” Primer Tm “Min= 57.0, Opt = 60.0, Max = 63.0.” PCR reactions were prepared in a 25 μL reaction volume containing: 1 μM forward and reverse primers, 0.1 nM dNTPs (NEB #N0447L), 1X Q5 buffer (NEB), 200–300 ng gDNA, 1 unit Q5 polymerase (NEB) and water. .. PCR reactions were conducted as follows: 95°C for 1 min, 30 cycles of 95°C for 10 s, melting temperature (Tm) for 10 s and 72°C for 45 s and a final step at 72°C for 1 min.

    Article Title: Using a soil bacterial species balance index to estimate potato crop productivity
    Article Snippet: .. Briefly, the gene specific sequence was fused to the Illumina TruSeq sequencing primers and PCR was carried out in a total volume of 50 μl that contained 1X Q5 buffer (NEB, Whitby, ON, Canada), 0.25 μM of each primer, 200 μM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase (NEB, Whitby, ON, Canada), and 1 μl of template DNA. ..

    Article Title: The hindgut microbiota of praying mantids is highly variable and includes both prey-associated and host-specific microbes
    Article Snippet: Library preparation and sequencing A previously described two-step PCR method was used to prepare the amplicon library for sequencing [ ]. .. The initial reaction contained 1X Q5 buffer (New England Biolabs [NEB], Ipswitch, MA), 200 μM dNTPS, 0.5 μM 515F, 0.5 μM 806R, 2 ng template DNA, and 0.02 U/μL Q5 Hot Start High-Fidelity DNA polymerase (NEB) for a total reaction volume of 10 μL.

    Article Title: Direct-fed microbial supplementation influences the bacteria community composition of the gastrointestinal tract of pre- and post-weaned calves
    Article Snippet: .. The PCR was carried out in a total volume of 50 µL that contains 1X Q5 buffer (NEB), 0.25 µM of each primer, 200 µM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase and 1 µL of template cDNA. .. The PCR started with an initial denaturation at 98 °C for 30 s followed by 10 cycles of denaturation at 98 °C for 10 s, annealing at 55 °C for 10 s and extension at 72 °C for 30 s, and 25 cycles of denaturation at 98 °C for 10 s, annealing at 65 °C for 10 s, extension at 72 °C for 30 s and a final extension step at 72 °C for 2 min.

    Article Title: Exacerbation induces a microbiota shift in sputa of COPD patients
    Article Snippet: .. PCR reactions contained: 1X Q5 buffer (New England Biolabs, Ipswich, Massachusetts, USA), 200 μM of each dNTP (Feldan, Quebec, Quebec, Canada), 0.2 μM of each 454 primer (IDT), 1 U of Q5 High-Fidelity DNA polymerase (NEB), and 1 μL of template DNA (10–50 ng). .. Cycling conditions were as follows: an initial denaturation at 98°C for 30 s, followed by 30 cycles of denaturation at 98°C for 10 s, annealing at 55°C for 30 s, extension at 72°C for 30 s, and a final extension at 72°C for 5 min. PCR products were purified using the Axyprep Mag PCR Clean-up Kit (Axygen, Union City, California, USA), and verified with a Bioanalyzer 2100 with DNA 7500 chips (Agilent Technologies, Santa Clara, California, USA).

    Article Title: The hindgut microbiota of praying mantids is highly variable and includes both prey-associated and host-specific microbes
    Article Snippet: PCR conditions were 98°C for 30 s for denaturation followed by 15 cycles at 98°C for 10 s, 52°C for 30 s, and 72°C for 30 s. The final extension step was 72°C for 2 min. .. The initial reaction product was added to the second reaction, which contained 1X Q5 buffer (NEB), 200 μM dNTPS, 0.5 μM 515F, 0.5 μM 806R, and 0.02 U/μL Hot Start High-Fidelity DNA polymerase (NEB) at a volume of 21 μL.

    Article Title: The Pokeweed Leaf mRNA Transcriptome and Its Regulation by Jasmonic Acid
    Article Snippet: .. Following cDNA synthesis, a PCR reaction was conducted, containing 1X Q5 buffer (NEB), 0.5 μM forward primer, 0.5 μM reverse primer, 200 mM dNTPs, 2 μL cDNA and 1 unit Q5 DNA polymerase (NEB) in a total volume of 50 μL. .. The PCR program included an initial denaturation of 95°C for 30 s, 30 cycles of 95°C for 30 s, 58°C for 30 s, 72°C for 120 s and finished with an extension at 72°C for 180 s. PCR products were separated on low-melt agarose and the correct size band excised and purified with EZ-10 Spin columns (Biobasic).

    DNA Sequencing:

    Article Title: CRISPR-Mediated Base Editing Enables Efficient Disruption of Eukaryotic Genes through Induction of STOP Codons
    Article Snippet: Paragraph title: RFLP assay and DNA sequencing ... The following genomic loci were targeted by sgSTOPs for SPRTN (chr1+ 231338561), SMARCAL1 (chr2+ 216414804), CHEK2 (chr22- 28734431), TIMELESS (chr12- 56431493) and FANCM ) using the default parameters with the following changes: Mispriming library = “HUMAN,” Primer size “min = 25, Opt = 27, Max = 30,” Primer Tm “Min= 57.0, Opt = 60.0, Max = 63.0.” PCR reactions were prepared in a 25 μL reaction volume containing: 1 μM forward and reverse primers, 0.1 nM dNTPs (NEB #N0447L), 1X Q5 buffer (NEB), 200–300 ng gDNA, 1 unit Q5 polymerase (NEB) and water.

    Sequencing:

    Article Title: Using a soil bacterial species balance index to estimate potato crop productivity
    Article Snippet: .. Briefly, the gene specific sequence was fused to the Illumina TruSeq sequencing primers and PCR was carried out in a total volume of 50 μl that contained 1X Q5 buffer (NEB, Whitby, ON, Canada), 0.25 μM of each primer, 200 μM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase (NEB, Whitby, ON, Canada), and 1 μl of template DNA. ..

    Article Title: The hindgut microbiota of praying mantids is highly variable and includes both prey-associated and host-specific microbes
    Article Snippet: Paragraph title: Library preparation and sequencing ... The initial reaction contained 1X Q5 buffer (New England Biolabs [NEB], Ipswitch, MA), 200 μM dNTPS, 0.5 μM 515F, 0.5 μM 806R, 2 ng template DNA, and 0.02 U/μL Q5 Hot Start High-Fidelity DNA polymerase (NEB) for a total reaction volume of 10 μL.

    Article Title: Direct-fed microbial supplementation influences the bacteria community composition of the gastrointestinal tract of pre- and post-weaned calves
    Article Snippet: Paragraph title: Amplification of bacterial ribosomal DNA and sequencing ... The PCR was carried out in a total volume of 50 µL that contains 1X Q5 buffer (NEB), 0.25 µM of each primer, 200 µM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase and 1 µL of template cDNA.

    Purification:

    Article Title: Using a soil bacterial species balance index to estimate potato crop productivity
    Article Snippet: Briefly, the gene specific sequence was fused to the Illumina TruSeq sequencing primers and PCR was carried out in a total volume of 50 μl that contained 1X Q5 buffer (NEB, Whitby, ON, Canada), 0.25 μM of each primer, 200 μM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase (NEB, Whitby, ON, Canada), and 1 μl of template DNA. .. The PCR reaction was purified using the Axygen PCR cleanup kit (Fisher Scientific, Nepean, ON, Canada).

    Article Title: Direct-fed microbial supplementation influences the bacteria community composition of the gastrointestinal tract of pre- and post-weaned calves
    Article Snippet: The PCR was carried out in a total volume of 50 µL that contains 1X Q5 buffer (NEB), 0.25 µM of each primer, 200 µM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase and 1 µL of template cDNA. .. The PCR reactions were purified using the Axygen PCR cleanup kit (Axygen).

    Article Title: Exacerbation induces a microbiota shift in sputa of COPD patients
    Article Snippet: PCR reactions contained: 1X Q5 buffer (New England Biolabs, Ipswich, Massachusetts, USA), 200 μM of each dNTP (Feldan, Quebec, Quebec, Canada), 0.2 μM of each 454 primer (IDT), 1 U of Q5 High-Fidelity DNA polymerase (NEB), and 1 μL of template DNA (10–50 ng). .. Cycling conditions were as follows: an initial denaturation at 98°C for 30 s, followed by 30 cycles of denaturation at 98°C for 10 s, annealing at 55°C for 30 s, extension at 72°C for 30 s, and a final extension at 72°C for 5 min. PCR products were purified using the Axyprep Mag PCR Clean-up Kit (Axygen, Union City, California, USA), and verified with a Bioanalyzer 2100 with DNA 7500 chips (Agilent Technologies, Santa Clara, California, USA).

    Article Title: The hindgut microbiota of praying mantids is highly variable and includes both prey-associated and host-specific microbes
    Article Snippet: The initial reaction product was added to the second reaction, which contained 1X Q5 buffer (NEB), 200 μM dNTPS, 0.5 μM 515F, 0.5 μM 806R, and 0.02 U/μL Hot Start High-Fidelity DNA polymerase (NEB) at a volume of 21 μL. .. Amplicons were purified using the EZ Cycle Pure Kit (Omega Biotek) before quantification with a NanoDrop Lite spectrophotometer (Thermo Scientific).

    Article Title: The Pokeweed Leaf mRNA Transcriptome and Its Regulation by Jasmonic Acid
    Article Snippet: Following cDNA synthesis, a PCR reaction was conducted, containing 1X Q5 buffer (NEB), 0.5 μM forward primer, 0.5 μM reverse primer, 200 mM dNTPs, 2 μL cDNA and 1 unit Q5 DNA polymerase (NEB) in a total volume of 50 μL. .. The PCR program included an initial denaturation of 95°C for 30 s, 30 cycles of 95°C for 30 s, 58°C for 30 s, 72°C for 120 s and finished with an extension at 72°C for 180 s. PCR products were separated on low-melt agarose and the correct size band excised and purified with EZ-10 Spin columns (Biobasic).

    Chromatin Immunoprecipitation:

    Article Title: Direct-fed microbial supplementation influences the bacteria community composition of the gastrointestinal tract of pre- and post-weaned calves
    Article Snippet: The PCR was carried out in a total volume of 50 µL that contains 1X Q5 buffer (NEB), 0.25 µM of each primer, 200 µM of each dNTPs, 1 U of Q5 High-Fidelity DNA polymerase and 1 µL of template cDNA. .. Quality of the purified PCR product was checked on a DNA7500 BioAnalyzer chip (Agilent) and quantified using a Nanodrop 1000 Spectrophotometer (Thermo Fisher Scientific).

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    New England Biolabs q5 reaction buffer
    Q5 Reaction Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 reaction buffer/product/New England Biolabs
    Average 99 stars, based on 19 article reviews
    Price from $9.99 to $1999.99
    q5 reaction buffer - by Bioz Stars, 2020-04
    99/100 stars
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