taq polymerase  (New England Biolabs)


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    Name:
    Taq DNA Polymerase with Standard Taq Buffer
    Description:
    Taq DNA Polymerase with Standard Taq Buffer 20 000 units
    Catalog Number:
    m0273e
    Price:
    1224
    Size:
    20 000 units
    Category:
    Thermostable DNA Polymerases
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    New England Biolabs taq polymerase
    Taq DNA Polymerase with Standard Taq Buffer
    Taq DNA Polymerase with Standard Taq Buffer 20 000 units
    https://www.bioz.com/result/taq polymerase/product/New England Biolabs
    Average 99 stars, based on 231 article reviews
    Price from $9.99 to $1999.99
    taq polymerase - by Bioz Stars, 2020-08
    99/100 stars

    Images

    1) Product Images from "PCR procedures to amplify GC-rich DNA sequences of Mycobacterium bovis"

    Article Title: PCR procedures to amplify GC-rich DNA sequences of Mycobacterium bovis

    Journal: bioRxiv

    doi: 10.1101/2020.02.18.953695

    Amplification results of three target genes using Taq polymerase. (A) Attempted amplification of the three DNA targets ( mpb83 , Mb012, and LMHCC_RS00060 ) sequences by Taq in the presence of DMSO using 3St, 2St and TD protocols; mpb83 and LMHCC_RS00060 successfully showed amplicons using the three protocols. Mb012 failed to show an amplicon. (B) Attempts of amplification of the three DNA target sequences in the absence of DMSO using 3St, 2St and TD protocols showing no amplicon. mpb83 with added enhancer in 3St PCR is used as a positive control. Mb0129 with no enhancer in 3St PCR is used as a negative control.
    Figure Legend Snippet: Amplification results of three target genes using Taq polymerase. (A) Attempted amplification of the three DNA targets ( mpb83 , Mb012, and LMHCC_RS00060 ) sequences by Taq in the presence of DMSO using 3St, 2St and TD protocols; mpb83 and LMHCC_RS00060 successfully showed amplicons using the three protocols. Mb012 failed to show an amplicon. (B) Attempts of amplification of the three DNA target sequences in the absence of DMSO using 3St, 2St and TD protocols showing no amplicon. mpb83 with added enhancer in 3St PCR is used as a positive control. Mb0129 with no enhancer in 3St PCR is used as a negative control.

    Techniques Used: Amplification, Polymerase Chain Reaction, Positive Control, Negative Control

    2) Product Images from "Sites of instability in the human TCF3 (E2A) gene adopt G-quadruplex DNA structures in vitro"

    Article Title: Sites of instability in the human TCF3 (E2A) gene adopt G-quadruplex DNA structures in vitro

    Journal: Frontiers in Genetics

    doi: 10.3389/fgene.2015.00177

    Guanine-rich templates from TCF3 and PBX1 block DNA synthesis in vitro . (A) Klenow polymerase extension assays using templates from the cytosine-rich strand (C-strand, left) or guanine-rich strand (G-strand, right) for each G4 sequence motif, resolved by denaturing PAGE. Reactions were performed in either KCl (K + ) or LiCl (Li + ). Shown are bands for stalled DNA synthesis (bracket) and full-length extension product (black arrow). G4 sequences begin at the bottom of the brackets. T-3′ differs from the motif listed in Table 1 , it includes 472 bp of surrounding genomic sequence (Supplementary Figure S2 ). (B) Primer extension reactions used Taq polymerase across a temperature range (50–80∘C) in either KCl (K + ) or (NH 4 ) 2 SO 4 (N) salt conditions on guanine-rich templates from TCF3 and PBX1 . Bands corresponding to stalled DNA synthesis (bracket) or full-length extension (black arrow) are shown.
    Figure Legend Snippet: Guanine-rich templates from TCF3 and PBX1 block DNA synthesis in vitro . (A) Klenow polymerase extension assays using templates from the cytosine-rich strand (C-strand, left) or guanine-rich strand (G-strand, right) for each G4 sequence motif, resolved by denaturing PAGE. Reactions were performed in either KCl (K + ) or LiCl (Li + ). Shown are bands for stalled DNA synthesis (bracket) and full-length extension product (black arrow). G4 sequences begin at the bottom of the brackets. T-3′ differs from the motif listed in Table 1 , it includes 472 bp of surrounding genomic sequence (Supplementary Figure S2 ). (B) Primer extension reactions used Taq polymerase across a temperature range (50–80∘C) in either KCl (K + ) or (NH 4 ) 2 SO 4 (N) salt conditions on guanine-rich templates from TCF3 and PBX1 . Bands corresponding to stalled DNA synthesis (bracket) or full-length extension (black arrow) are shown.

    Techniques Used: Blocking Assay, DNA Synthesis, In Vitro, Sequencing, Polyacrylamide Gel Electrophoresis

    3) Product Images from "Antibody-Directed Lentiviral Gene Transduction for Live-Cell Monitoring and Selection of Human iPS and hES Cells"

    Article Title: Antibody-Directed Lentiviral Gene Transduction for Live-Cell Monitoring and Selection of Human iPS and hES Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0034778

    Characterization of endogenous pluripotent makers in selected iPS cell lines. Panel A. Total RNA was isolated using RNeasy Micro Kit from selected iPS cell lines (G1–G3, G5, G6), hES H9 cells (H9), and human primary fibroblasts (F). Total RNA (500 ng) was reverse-transcribed using Superscript III Reverse Transcriptase primed with oligo(dT) 12–18 and used as template in subsequent PCR with Taq DNA Polymerase. PCR analysis examined the expression of endogenous Oct4, Nanog, Sox2, as well as ABCG2, Rex1, DNMT3B and hTERT. GAPDH was used as an internal control. N, no template control (N). PCR products were analyzed on a 10% polyacrylamide TBE Precast Gel. Panel B. TRAP assay for telomerase activity. Selected iPS cells (G1–G3, G6), hES H9 cells (H9), and human primary fibroblasts (F) were analyzed for telomerase activity using the TRAPEZE RT Telomerase Detection Kit as described in M M. PCR products were separated on 10% polyacrylamide TBE Precast Gel. Individual samples are as indicated.
    Figure Legend Snippet: Characterization of endogenous pluripotent makers in selected iPS cell lines. Panel A. Total RNA was isolated using RNeasy Micro Kit from selected iPS cell lines (G1–G3, G5, G6), hES H9 cells (H9), and human primary fibroblasts (F). Total RNA (500 ng) was reverse-transcribed using Superscript III Reverse Transcriptase primed with oligo(dT) 12–18 and used as template in subsequent PCR with Taq DNA Polymerase. PCR analysis examined the expression of endogenous Oct4, Nanog, Sox2, as well as ABCG2, Rex1, DNMT3B and hTERT. GAPDH was used as an internal control. N, no template control (N). PCR products were analyzed on a 10% polyacrylamide TBE Precast Gel. Panel B. TRAP assay for telomerase activity. Selected iPS cells (G1–G3, G6), hES H9 cells (H9), and human primary fibroblasts (F) were analyzed for telomerase activity using the TRAPEZE RT Telomerase Detection Kit as described in M M. PCR products were separated on 10% polyacrylamide TBE Precast Gel. Individual samples are as indicated.

    Techniques Used: Isolation, Polymerase Chain Reaction, Expressing, TRAP Assay, Activity Assay

    4) Product Images from "Next-generation mapping: a novel approach for detection of pathogenic structural variants with a potential utility in clinical diagnosis"

    Article Title: Next-generation mapping: a novel approach for detection of pathogenic structural variants with a potential utility in clinical diagnosis

    Journal: Genome Medicine

    doi: 10.1186/s13073-017-0479-0

    DNA labeling for NGM. The DNA labeling workflow is divided into four consecutive steps. First, the high molecular weight DNA is nicked with an endonuclease of choice that introduces single strand nicks throughout the genome. Second, Taq polymerase recognizes these sites and replaces several nucleotides with fluorescently tagged nucleotides added to the solution. Third, the two ends of the DNA are ligated together using DNA ligase. Fourth, the DNA backbone is stained with DNA Stain
    Figure Legend Snippet: DNA labeling for NGM. The DNA labeling workflow is divided into four consecutive steps. First, the high molecular weight DNA is nicked with an endonuclease of choice that introduces single strand nicks throughout the genome. Second, Taq polymerase recognizes these sites and replaces several nucleotides with fluorescently tagged nucleotides added to the solution. Third, the two ends of the DNA are ligated together using DNA ligase. Fourth, the DNA backbone is stained with DNA Stain

    Techniques Used: DNA Labeling, Molecular Weight, Staining

    5) Product Images from "Direct Chromatin PCR (DC-PCR): Hypotonic Conditions Allow Differentiation of Chromatin States during Thermal Cycling"

    Article Title: Direct Chromatin PCR (DC-PCR): Hypotonic Conditions Allow Differentiation of Chromatin States during Thermal Cycling

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0044690

    Direct Amplification of open chromatin with phi29 polymerase: Translation of DC-PCR to genome-wide chromatin analysis. CDKN2A PCR on KMS-12-PE supernatant after direct genome-wide phi29 amplification ( A ). Untreated KMS-12-PE cells or cells treated with DAC (1 µM for 3 d) were placed directly in phi29 reaction mix for 4.5 hrs at 30°C. After that, the reaction was centrifuged to pellet cell carcasses. Only the upper 50% of the supernatant was used for subsequent amplification of CDKN2A sites by conventional PCR. Supernatant from DAC treated cells yielded more CDKN2A products than vehicle treated cells. When DAC treated cells were incubated in reaction buffer without Phi29 polymerase no CDKN2A products could be amplified from supernatants by taq polymerase during conventional PCR. FOSB PCR on KMS-12-PE supernatants obtained from untreated cells or cells treated with 10-E-09 (10 µM for 6 h) after direct genome-wide phi29 amplification ( B ). The same procedure as above yielded FOSB PCR products from supernatants of 10-E-09 treated KMS-12-PE cells amplified by phi29 while vehicle treated cells or supernatants from 10-E-09 treated cells incubated in reaction buffer without phi29 yielded no FOSB regulatory region amplicons. Results are representative of three independent experiments.
    Figure Legend Snippet: Direct Amplification of open chromatin with phi29 polymerase: Translation of DC-PCR to genome-wide chromatin analysis. CDKN2A PCR on KMS-12-PE supernatant after direct genome-wide phi29 amplification ( A ). Untreated KMS-12-PE cells or cells treated with DAC (1 µM for 3 d) were placed directly in phi29 reaction mix for 4.5 hrs at 30°C. After that, the reaction was centrifuged to pellet cell carcasses. Only the upper 50% of the supernatant was used for subsequent amplification of CDKN2A sites by conventional PCR. Supernatant from DAC treated cells yielded more CDKN2A products than vehicle treated cells. When DAC treated cells were incubated in reaction buffer without Phi29 polymerase no CDKN2A products could be amplified from supernatants by taq polymerase during conventional PCR. FOSB PCR on KMS-12-PE supernatants obtained from untreated cells or cells treated with 10-E-09 (10 µM for 6 h) after direct genome-wide phi29 amplification ( B ). The same procedure as above yielded FOSB PCR products from supernatants of 10-E-09 treated KMS-12-PE cells amplified by phi29 while vehicle treated cells or supernatants from 10-E-09 treated cells incubated in reaction buffer without phi29 yielded no FOSB regulatory region amplicons. Results are representative of three independent experiments.

    Techniques Used: Amplification, Polymerase Chain Reaction, Genome Wide, Incubation

    6) Product Images from "High-resolution mapping of DNA polymerase fidelity using nucleotide imbalances and next-generation sequencing"

    Article Title: High-resolution mapping of DNA polymerase fidelity using nucleotide imbalances and next-generation sequencing

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gky296

    DNA polymerase error preferences. Heat maps reflect nucleotide substitution and single nucleotide deletion preferences of ( A ) Dpo4, ( B ) Taq, ( C ) Sequenase 2.0, ( D ) AMV RT and ( E ) Phi29 copying in ‘T’, ‘A’, ‘C’ and ‘G’ template contexts. Error fraction was determined by normalizing individual error subtype frequencies to the total error rate measured at the lowest [dRTP] tested (10 −7 μM) for each template context. Values are the average of two experiments.
    Figure Legend Snippet: DNA polymerase error preferences. Heat maps reflect nucleotide substitution and single nucleotide deletion preferences of ( A ) Dpo4, ( B ) Taq, ( C ) Sequenase 2.0, ( D ) AMV RT and ( E ) Phi29 copying in ‘T’, ‘A’, ‘C’ and ‘G’ template contexts. Error fraction was determined by normalizing individual error subtype frequencies to the total error rate measured at the lowest [dRTP] tested (10 −7 μM) for each template context. Values are the average of two experiments.

    Techniques Used:

    MagNIFi assay validation. ( A ) Fidelity dose response curves of Sequenase 2.0, AMV RT, Phi29, Dpo4 and Taq copying in ‘T’ template contexts. Values are the average of two experiments. Standard deviation error bars ( n = 2) are smaller than data points. Curves show qualitative agreement between DNA polymerase FC 50 values (indicated by black dotted line) and the expected rank order of natural DNA polymerase error rates ( Supplementary Table S4 ). In general, fidelity increases from right to left. ( B ) Calibration between error rate and [FC 50 ]. We show a calibration curve relating multiple reported error rates per DNA polymerase ( Supplementary Table S4 ) and the average FC 50 value of each DNA polymerase ( Supplementary Table S3 ). Nonlinear fitting on a log–log plot (line of best fit in grey) revealed the following equation: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} }{}$y = 10^{(2.063log(x) + 1.557)}$\end{document} , RMSE = 0.0008998 errors/bp.
    Figure Legend Snippet: MagNIFi assay validation. ( A ) Fidelity dose response curves of Sequenase 2.0, AMV RT, Phi29, Dpo4 and Taq copying in ‘T’ template contexts. Values are the average of two experiments. Standard deviation error bars ( n = 2) are smaller than data points. Curves show qualitative agreement between DNA polymerase FC 50 values (indicated by black dotted line) and the expected rank order of natural DNA polymerase error rates ( Supplementary Table S4 ). In general, fidelity increases from right to left. ( B ) Calibration between error rate and [FC 50 ]. We show a calibration curve relating multiple reported error rates per DNA polymerase ( Supplementary Table S4 ) and the average FC 50 value of each DNA polymerase ( Supplementary Table S3 ). Nonlinear fitting on a log–log plot (line of best fit in grey) revealed the following equation: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} }{}$y = 10^{(2.063log(x) + 1.557)}$\end{document} , RMSE = 0.0008998 errors/bp.

    Techniques Used: Standard Deviation

    7) Product Images from "High-resolution mapping of DNA polymerase fidelity using nucleotide imbalances and next-generation sequencing"

    Article Title: High-resolution mapping of DNA polymerase fidelity using nucleotide imbalances and next-generation sequencing

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gky296

    DNA polymerase error preferences. Heat maps reflect nucleotide substitution and single nucleotide deletion preferences of ( A ) Dpo4, ( B ) Taq, ( C ) Sequenase 2.0, ( D ) AMV RT and ( E ) Phi29 copying in ‘T’, ‘A’, ‘C’ and ‘G’ template contexts. Error fraction was determined by normalizing individual error subtype frequencies to the total error rate measured at the lowest [dRTP] tested (10 −7 μM) for each template context. Values are the average of two experiments.
    Figure Legend Snippet: DNA polymerase error preferences. Heat maps reflect nucleotide substitution and single nucleotide deletion preferences of ( A ) Dpo4, ( B ) Taq, ( C ) Sequenase 2.0, ( D ) AMV RT and ( E ) Phi29 copying in ‘T’, ‘A’, ‘C’ and ‘G’ template contexts. Error fraction was determined by normalizing individual error subtype frequencies to the total error rate measured at the lowest [dRTP] tested (10 −7 μM) for each template context. Values are the average of two experiments.

    Techniques Used:

    MagNIFi assay validation. ( A ) Fidelity dose response curves of Sequenase 2.0, AMV RT, Phi29, Dpo4 and Taq copying in ‘T’ template contexts. Values are the average of two experiments. Standard deviation error bars ( n = 2) are smaller than data points. Curves show qualitative agreement between DNA polymerase FC 50 ). In general, fidelity increases from right to left. ( B ) Calibration between error rate and [FC 50 ) and the average FC 50 ). Nonlinear fitting on a log–log plot (line of best fit in grey) revealed the following equation: , RMSE = 0.0008998 errors/bp.
    Figure Legend Snippet: MagNIFi assay validation. ( A ) Fidelity dose response curves of Sequenase 2.0, AMV RT, Phi29, Dpo4 and Taq copying in ‘T’ template contexts. Values are the average of two experiments. Standard deviation error bars ( n = 2) are smaller than data points. Curves show qualitative agreement between DNA polymerase FC 50 ). In general, fidelity increases from right to left. ( B ) Calibration between error rate and [FC 50 ) and the average FC 50 ). Nonlinear fitting on a log–log plot (line of best fit in grey) revealed the following equation: , RMSE = 0.0008998 errors/bp.

    Techniques Used: Standard Deviation

    8) Product Images from "In situ 10-cell RNA sequencing in tissue and tumor biopsy samples"

    Article Title: In situ 10-cell RNA sequencing in tissue and tumor biopsy samples

    Journal: Scientific Reports

    doi: 10.1038/s41598-019-41235-9

    A blend of Taq–Phusion polymerases improves selective poly(A) amplification of cDNA and reduces AL1 primer requirements. Cells were obtained by LCM from a human breast biopsy and split into 10-cell equivalent amplification replicates. ( A ) Poly(A) PCR was performed with 15 µg of AL1 primer with Taq alone (10 units), Phusion alone (4 units) or Taq/Phusion combination (3.75 units/1.5 units). ( B ) Poly(A) PCR was performed with either 25, 5, 2.5, or 0.5 µg of AL1 primer and the Taq–Phusion blend from (A). Above—Relative abundance for the indicated genes and preamplification conditions was measured by quantitative PCR (qPCR). Data are shown as the median inverse quantification cycle (40–Cq) ± range from n = 3 amplification replicates and were analysed by two-way (A) or one-way (B) ANOVA with replication. Below—Preamplifications were analysed by agarose gel electrophoresis to separate poly(A)-amplified cDNA from nonspecific, low molecular-weight concatemer (n.s.). Qualitatively similar results were obtained separately three times. Lanes were cropped by poly(A) PCR cycles for display but were electrophoresed on the same agarose gel and processed identically. The uncropped image is shown in Supplementary Fig. S13A .
    Figure Legend Snippet: A blend of Taq–Phusion polymerases improves selective poly(A) amplification of cDNA and reduces AL1 primer requirements. Cells were obtained by LCM from a human breast biopsy and split into 10-cell equivalent amplification replicates. ( A ) Poly(A) PCR was performed with 15 µg of AL1 primer with Taq alone (10 units), Phusion alone (4 units) or Taq/Phusion combination (3.75 units/1.5 units). ( B ) Poly(A) PCR was performed with either 25, 5, 2.5, or 0.5 µg of AL1 primer and the Taq–Phusion blend from (A). Above—Relative abundance for the indicated genes and preamplification conditions was measured by quantitative PCR (qPCR). Data are shown as the median inverse quantification cycle (40–Cq) ± range from n = 3 amplification replicates and were analysed by two-way (A) or one-way (B) ANOVA with replication. Below—Preamplifications were analysed by agarose gel electrophoresis to separate poly(A)-amplified cDNA from nonspecific, low molecular-weight concatemer (n.s.). Qualitatively similar results were obtained separately three times. Lanes were cropped by poly(A) PCR cycles for display but were electrophoresed on the same agarose gel and processed identically. The uncropped image is shown in Supplementary Fig. S13A .

    Techniques Used: Amplification, Laser Capture Microdissection, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Molecular Weight

    Related Articles

    Nucleic Acid Electrophoresis:

    Article Title: A one-tube method for rapid and reliable plant genomic DNA isolation for PCR analysis
    Article Snippet: .. PCR amplification and gel electrophoresis For routine PCR, 1 ∼ 1.5 μl of DNA extract was added to 20 μl of PCR reaction mix containing standard PCR buffer and 1 unit of Taq DNA polymerase (Cat # = M0273, New England Biolabs, Beverly, MA). .. For long-range PCR, 1.5 μl of DNA extract was added to 25 μl of LongAmp Taq DNA polymerase PCR reaction mix (Cat # = M0323, New England Biolabs) following the manufacturer’s protocol.

    Amplification:

    Article Title: A one-tube method for rapid and reliable plant genomic DNA isolation for PCR analysis
    Article Snippet: .. PCR amplification and gel electrophoresis For routine PCR, 1 ∼ 1.5 μl of DNA extract was added to 20 μl of PCR reaction mix containing standard PCR buffer and 1 unit of Taq DNA polymerase (Cat # = M0273, New England Biolabs, Beverly, MA). .. For long-range PCR, 1.5 μl of DNA extract was added to 25 μl of LongAmp Taq DNA polymerase PCR reaction mix (Cat # = M0323, New England Biolabs) following the manufacturer’s protocol.

    Article Title: Enzymatic Cleavage of 3’-Esterified Nucleotides Enables a Long, Continuous DNA Synthesis
    Article Snippet: .. The amplified PCR products were A-tailed by Taq DNA polymerase (NEB). .. The A-tailing products were purified by Qiagen PCR clean-up kit, and then ligated with the TA cloning vector (Yeastern Biotech, Taipei, Taiwan) following the manufacturer’s protocol.

    Article Title: One enzyme reverse transcription qPCR using Taq DNA polymerase
    Article Snippet: .. We hypothesized that the buffers in which Taq DNA polymerase is commonly used have been optimized for DNA amplification and likely would not support robust reverse transcription. ..

    Labeling:

    Article Title: CRISPR-CAS9 D10A nickase target-specific fluorescent labeling of double strand DNA for whole genome mapping and structural variation analysis
    Article Snippet: .. The two color genome mapping with Cas9n fluorescent nick-labeling and sequence-motif labeling After nicking with Cas9n D10A as previously described in the Cas9n fluorescent nick-labeling section, the sample was digested with RNAseA (190 ng/μL, QIAGEN) at 37°C for 20 min. After digestion, the sample was labeled with ATTO 532-dATP, dTGC (100 nM) and 2.5 units of DNA Taq Polymerase (NEB) in the presence of 1X Thermopol Buffer (NEB) at 72°C for 1 h. The sample was treated with 1 unit of SAP (USB Products) and RNAseA (100 ng/μL) at 37°C for 20 min and then 65°C for 15 min. ..

    Article Title: Sites of instability in the human TCF3 (E2A) gene adopt G-quadruplex DNA structures in vitro
    Article Snippet: .. Single-stranded phagemid templates were primed with a 32 P 5′ end labeled M13 forward primer, which was extended with Klenow or Taq polymerase (NEB). .. In Klenow reactions, KCl or LiCl was added to a final concentration of 25 mM.

    Polymerase Chain Reaction:

    Article Title: PCR procedures to amplify GC-rich DNA sequences of Mycobacterium bovis
    Article Snippet: .. Unsuccessful attempts were faced with Taq polymerase, OneTaq DNA Polymerase (NEB, M0480S), Platinum™ Pfx DNA Polymerase (Invitrogen, 11708039), Expand Long Template PCR System (an enzyme mix that contains thermostable Taq DNA Polymerase and a thermostable DNA polymerase with proofreading activity, Roche, 11681834001). .. This observation prompted us to establish a reliable PCR procedure that not only can be used for cloning the M. bovis sequences but also can be applied to the amplification other targets of high GC content.

    Article Title: A one-tube method for rapid and reliable plant genomic DNA isolation for PCR analysis
    Article Snippet: .. PCR amplification and gel electrophoresis For routine PCR, 1 ∼ 1.5 μl of DNA extract was added to 20 μl of PCR reaction mix containing standard PCR buffer and 1 unit of Taq DNA polymerase (Cat # = M0273, New England Biolabs, Beverly, MA). .. For long-range PCR, 1.5 μl of DNA extract was added to 25 μl of LongAmp Taq DNA polymerase PCR reaction mix (Cat # = M0323, New England Biolabs) following the manufacturer’s protocol.

    Article Title: Enzymatic Cleavage of 3’-Esterified Nucleotides Enables a Long, Continuous DNA Synthesis
    Article Snippet: .. The amplified PCR products were A-tailed by Taq DNA polymerase (NEB). .. The A-tailing products were purified by Qiagen PCR clean-up kit, and then ligated with the TA cloning vector (Yeastern Biotech, Taipei, Taiwan) following the manufacturer’s protocol.

    Activity Assay:

    Article Title: PCR procedures to amplify GC-rich DNA sequences of Mycobacterium bovis
    Article Snippet: .. Unsuccessful attempts were faced with Taq polymerase, OneTaq DNA Polymerase (NEB, M0480S), Platinum™ Pfx DNA Polymerase (Invitrogen, 11708039), Expand Long Template PCR System (an enzyme mix that contains thermostable Taq DNA Polymerase and a thermostable DNA polymerase with proofreading activity, Roche, 11681834001). .. This observation prompted us to establish a reliable PCR procedure that not only can be used for cloning the M. bovis sequences but also can be applied to the amplification other targets of high GC content.

    Quantitative RT-PCR:

    Article Title: One enzyme reverse transcription qPCR using Taq DNA polymerase
    Article Snippet: .. These results suggest that Taq DNA polymerase can support TaqMan RT-qPCR analyses of RNA in one-enzyme reactions. .. To further prove that the reverse transcriptase is inherent in Taq polymerase itself, we incubated RT-qPCR assays at 95 °C for 5 min prior to reverse transcription, which should inactivate any contaminating mesophilic reverse transcriptases ( Supplementary Figure 1 ).

    Sequencing:

    Article Title: CRISPR-CAS9 D10A nickase target-specific fluorescent labeling of double strand DNA for whole genome mapping and structural variation analysis
    Article Snippet: .. The two color genome mapping with Cas9n fluorescent nick-labeling and sequence-motif labeling After nicking with Cas9n D10A as previously described in the Cas9n fluorescent nick-labeling section, the sample was digested with RNAseA (190 ng/μL, QIAGEN) at 37°C for 20 min. After digestion, the sample was labeled with ATTO 532-dATP, dTGC (100 nM) and 2.5 units of DNA Taq Polymerase (NEB) in the presence of 1X Thermopol Buffer (NEB) at 72°C for 1 h. The sample was treated with 1 unit of SAP (USB Products) and RNAseA (100 ng/μL) at 37°C for 20 min and then 65°C for 15 min. ..

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    New England Biolabs longamp taq dna polymerase
    Thermal shock is crucial for successful PCR amplification from fungal spores. A. fumigatus spores at three different concentrations (8 × 10 8 /ml, 1 × 10 8 /ml, and 5 × 10 7 /ml) were prepared as described in Basic Protocol 1 . 3 µl of spore suspension were used as the <t>DNA</t> template for PCR. ( A ) PCR result from spore suspension subjected to thermal shock of 95°C for 15 min and −80°C for 10 min prior to PCR, as described in Basic Protocol 2 . No thermal shock was done for spore suspensions from Panels B and C . Subsequently, a PCR was run with an initial denaturation step of 95°C for 1 min (A), 5 min (B), or 15 min (C) with primers ITS1 and D2 (expected PCR product ∼1.2 kb) and <t>LongAmp</t> <t>Taq</t> DNA polymerase with PCR conditions described in Basic Protocol 2 . P: positive PCR control amplified from genomic DNA (50 ng) of the A. fumigatus wild‐type strain; N: negative control (no DNA).
    Longamp Taq Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/longamp taq dna polymerase/product/New England Biolabs
    Average 99 stars, based on 140 article reviews
    Price from $9.99 to $1999.99
    longamp taq dna polymerase - by Bioz Stars, 2020-08
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    85
    New England Biolabs target dna fragments taq dna polymerase
    Robust PCR-amplification of insert <t>DNA</t> fragments using deoxyinosine-containing primers. Analytical agarose gel electrophoresis of PCR products produced by <t>Taq</t> polymerase using either plasmid DNA (A) or E. coli colonies (B) as template material. Relative to the calculated T m , annealing temperatures used for PCR cycling are indicated for each lane.
    Target Dna Fragments Taq Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/target dna fragments taq dna polymerase/product/New England Biolabs
    Average 85 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
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    New England Biolabs taq polymerase
    Amplification results of three target genes using <t>Taq</t> polymerase. (A) Attempted amplification of the three <t>DNA</t> targets ( mpb83 , Mb012, and LMHCC_RS00060 ) sequences by Taq in the presence of DMSO using 3St, 2St and TD protocols; mpb83 and LMHCC_RS00060 successfully showed amplicons using the three protocols. Mb012 failed to show an amplicon. (B) Attempts of amplification of the three DNA target sequences in the absence of DMSO using 3St, 2St and TD protocols showing no amplicon. mpb83 with added enhancer in 3St PCR is used as a positive control. Mb0129 with no enhancer in 3St PCR is used as a negative control.
    Taq Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 346 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taq polymerase/product/New England Biolabs
    Average 99 stars, based on 346 article reviews
    Price from $9.99 to $1999.99
    taq polymerase - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

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    Thermal shock is crucial for successful PCR amplification from fungal spores. A. fumigatus spores at three different concentrations (8 × 10 8 /ml, 1 × 10 8 /ml, and 5 × 10 7 /ml) were prepared as described in Basic Protocol 1 . 3 µl of spore suspension were used as the DNA template for PCR. ( A ) PCR result from spore suspension subjected to thermal shock of 95°C for 15 min and −80°C for 10 min prior to PCR, as described in Basic Protocol 2 . No thermal shock was done for spore suspensions from Panels B and C . Subsequently, a PCR was run with an initial denaturation step of 95°C for 1 min (A), 5 min (B), or 15 min (C) with primers ITS1 and D2 (expected PCR product ∼1.2 kb) and LongAmp Taq DNA polymerase with PCR conditions described in Basic Protocol 2 . P: positive PCR control amplified from genomic DNA (50 ng) of the A. fumigatus wild‐type strain; N: negative control (no DNA).

    Journal: Current Protocols in Microbiology

    Article Title: Fast and Reliable PCR Amplification from Aspergillus fumigatus Spore Suspension Without Traditional DNA Extraction). Fast and reliable PCR amplification from Aspergillus fumigatus spore suspension without traditional DNA extraction

    doi: 10.1002/cpmc.89

    Figure Lengend Snippet: Thermal shock is crucial for successful PCR amplification from fungal spores. A. fumigatus spores at three different concentrations (8 × 10 8 /ml, 1 × 10 8 /ml, and 5 × 10 7 /ml) were prepared as described in Basic Protocol 1 . 3 µl of spore suspension were used as the DNA template for PCR. ( A ) PCR result from spore suspension subjected to thermal shock of 95°C for 15 min and −80°C for 10 min prior to PCR, as described in Basic Protocol 2 . No thermal shock was done for spore suspensions from Panels B and C . Subsequently, a PCR was run with an initial denaturation step of 95°C for 1 min (A), 5 min (B), or 15 min (C) with primers ITS1 and D2 (expected PCR product ∼1.2 kb) and LongAmp Taq DNA polymerase with PCR conditions described in Basic Protocol 2 . P: positive PCR control amplified from genomic DNA (50 ng) of the A. fumigatus wild‐type strain; N: negative control (no DNA).

    Article Snippet: Materials Spore suspension (Basic Protocol ) Primers (5 µM each, forward and reverse; experiment specific) LongAmp Taq DNA polymerase with 5× reaction buffer (New England Biolabs; M0323) dNTP mix (5 mM of each dNTP) Sterile, molecular‐grade water For agarose gel electrophoresis (also see Current Protocols article: Voytas, ): 6× DNA loading dye 1× TAE buffer (see recipe) Ethidium bromide or other DNA gel stain 0.2‐ml PCR tubes, 0.2‐ml/well 96‐well PCR plates, or 150 µl/well 384 well PCR plates with tight (preferably aluminum) seals Thermal cycler Centrifuge UV transilluminator Additional reagents and equipment for agarose gel electrophoresis (see Current Protocols article: Voytas, )

    Techniques: Polymerase Chain Reaction, Amplification, Negative Control

    Spore PCR using the supernatant from spore suspensions of different filamentous fungi with primers ITS1/D2 (expected PCR band size is ∼1.2 kb) and ITS1/ITS4 (expected PCR band size ∼600 bp). Two different spore concentrations were tested (i.e., 5 × 10 7 /ml and 1 × 10 7 /ml). 1 µl of the supernatant was used in the PCR reaction with the LongAmp Taq DNA polymerase. Positive PCR controls were amplified from DNA (50 ng) of the A. fumigatus wild‐type strain with primers ITS1/D2 (P1) and ITS1/ITS4 (P2). N: negative control (no DNA).

    Journal: Current Protocols in Microbiology

    Article Title: Fast and Reliable PCR Amplification from Aspergillus fumigatus Spore Suspension Without Traditional DNA Extraction). Fast and reliable PCR amplification from Aspergillus fumigatus spore suspension without traditional DNA extraction

    doi: 10.1002/cpmc.89

    Figure Lengend Snippet: Spore PCR using the supernatant from spore suspensions of different filamentous fungi with primers ITS1/D2 (expected PCR band size is ∼1.2 kb) and ITS1/ITS4 (expected PCR band size ∼600 bp). Two different spore concentrations were tested (i.e., 5 × 10 7 /ml and 1 × 10 7 /ml). 1 µl of the supernatant was used in the PCR reaction with the LongAmp Taq DNA polymerase. Positive PCR controls were amplified from DNA (50 ng) of the A. fumigatus wild‐type strain with primers ITS1/D2 (P1) and ITS1/ITS4 (P2). N: negative control (no DNA).

    Article Snippet: Materials Spore suspension (Basic Protocol ) Primers (5 µM each, forward and reverse; experiment specific) LongAmp Taq DNA polymerase with 5× reaction buffer (New England Biolabs; M0323) dNTP mix (5 mM of each dNTP) Sterile, molecular‐grade water For agarose gel electrophoresis (also see Current Protocols article: Voytas, ): 6× DNA loading dye 1× TAE buffer (see recipe) Ethidium bromide or other DNA gel stain 0.2‐ml PCR tubes, 0.2‐ml/well 96‐well PCR plates, or 150 µl/well 384 well PCR plates with tight (preferably aluminum) seals Thermal cycler Centrifuge UV transilluminator Additional reagents and equipment for agarose gel electrophoresis (see Current Protocols article: Voytas, )

    Techniques: Polymerase Chain Reaction, Amplification, Negative Control

    Robust PCR-amplification of insert DNA fragments using deoxyinosine-containing primers. Analytical agarose gel electrophoresis of PCR products produced by Taq polymerase using either plasmid DNA (A) or E. coli colonies (B) as template material. Relative to the calculated T m , annealing temperatures used for PCR cycling are indicated for each lane.

    Journal: BMC Biotechnology

    Article Title: Directional cloning of DNA fragments using deoxyinosine-containing oligonucleotides and endonuclease V

    doi: 10.1186/1472-6750-13-81

    Figure Lengend Snippet: Robust PCR-amplification of insert DNA fragments using deoxyinosine-containing primers. Analytical agarose gel electrophoresis of PCR products produced by Taq polymerase using either plasmid DNA (A) or E. coli colonies (B) as template material. Relative to the calculated T m , annealing temperatures used for PCR cycling are indicated for each lane.

    Article Snippet: PCR-based amplification of target DNA fragments Taq DNA polymerase and dNTP mix were obtained from New England Biolabs (Frankfurt am Main, Germany).

    Techniques: Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Produced, Plasmid Preparation

    Amplification results of three target genes using Taq polymerase. (A) Attempted amplification of the three DNA targets ( mpb83 , Mb012, and LMHCC_RS00060 ) sequences by Taq in the presence of DMSO using 3St, 2St and TD protocols; mpb83 and LMHCC_RS00060 successfully showed amplicons using the three protocols. Mb012 failed to show an amplicon. (B) Attempts of amplification of the three DNA target sequences in the absence of DMSO using 3St, 2St and TD protocols showing no amplicon. mpb83 with added enhancer in 3St PCR is used as a positive control. Mb0129 with no enhancer in 3St PCR is used as a negative control.

    Journal: bioRxiv

    Article Title: PCR procedures to amplify GC-rich DNA sequences of Mycobacterium bovis

    doi: 10.1101/2020.02.18.953695

    Figure Lengend Snippet: Amplification results of three target genes using Taq polymerase. (A) Attempted amplification of the three DNA targets ( mpb83 , Mb012, and LMHCC_RS00060 ) sequences by Taq in the presence of DMSO using 3St, 2St and TD protocols; mpb83 and LMHCC_RS00060 successfully showed amplicons using the three protocols. Mb012 failed to show an amplicon. (B) Attempts of amplification of the three DNA target sequences in the absence of DMSO using 3St, 2St and TD protocols showing no amplicon. mpb83 with added enhancer in 3St PCR is used as a positive control. Mb0129 with no enhancer in 3St PCR is used as a negative control.

    Article Snippet: Unsuccessful attempts were faced with Taq polymerase, OneTaq DNA Polymerase (NEB, M0480S), Platinum™ Pfx DNA Polymerase (Invitrogen, 11708039), Expand Long Template PCR System (an enzyme mix that contains thermostable Taq DNA Polymerase and a thermostable DNA polymerase with proofreading activity, Roche, 11681834001).

    Techniques: Amplification, Polymerase Chain Reaction, Positive Control, Negative Control

    Effect of DNase I treatment on Taq DNA polymerase-mediated RT-qPCR assay. Taq DNA polymerase purchased from NEB was used to operate CDC SARS-CoV-2 N1, N2, and N3 TaqMan RT-qPCR assays using SARS-CoV-2 viral genomic RNA (panels A-C) or N gene armored RNA (panels D-F) treated with DNase I. Amplification curves shown in panels A-C resulted from 6000 (black traces), 600 (red traces), 60 (blue traces), 6 (pink traces), and 0 (gray traces) copies of SARS-CoV-2 genomic RNA. Amplification curves in panels D-F resulted from 30,000 (black traces), 3,000 (red traces), 300 (blue traces), 30 (pink traces) and 0 (gray traces) copies of N gene armored RNA. Representative Ct values for RT-qPCR amplification of indicated copies of untreated and DNase I treated SARS-CoV-2 genomic RNA and N gene armored RNA are tabulated.

    Journal: bioRxiv

    Article Title: One enzyme reverse transcription qPCR using Taq DNA polymerase

    doi: 10.1101/2020.05.27.120238

    Figure Lengend Snippet: Effect of DNase I treatment on Taq DNA polymerase-mediated RT-qPCR assay. Taq DNA polymerase purchased from NEB was used to operate CDC SARS-CoV-2 N1, N2, and N3 TaqMan RT-qPCR assays using SARS-CoV-2 viral genomic RNA (panels A-C) or N gene armored RNA (panels D-F) treated with DNase I. Amplification curves shown in panels A-C resulted from 6000 (black traces), 600 (red traces), 60 (blue traces), 6 (pink traces), and 0 (gray traces) copies of SARS-CoV-2 genomic RNA. Amplification curves in panels D-F resulted from 30,000 (black traces), 3,000 (red traces), 300 (blue traces), 30 (pink traces) and 0 (gray traces) copies of N gene armored RNA. Representative Ct values for RT-qPCR amplification of indicated copies of untreated and DNase I treated SARS-CoV-2 genomic RNA and N gene armored RNA are tabulated.

    Article Snippet: These results suggest that Taq DNA polymerase can support TaqMan RT-qPCR analyses of RNA in one-enzyme reactions.

    Techniques: Quantitative RT-PCR, Amplification

    SARS-CoV-2 N1 TaqMan RT-qPCR assays performed using NEB Taq DNA polymerase and N gene armored RNA in indicated buffers. Buffer compositions are detailed in Table 2 . Amplification curves resulting from 3 × 10 5 (black traces), 3 × 10 4 (red traces), 3 × 10 3 (blue traces), 3 × 10 2 (pink traces), 30 (green traces), and 0 (gray) copies of SARS-CoV-2 N gene armored RNA are depicted.

    Journal: bioRxiv

    Article Title: One enzyme reverse transcription qPCR using Taq DNA polymerase

    doi: 10.1101/2020.05.27.120238

    Figure Lengend Snippet: SARS-CoV-2 N1 TaqMan RT-qPCR assays performed using NEB Taq DNA polymerase and N gene armored RNA in indicated buffers. Buffer compositions are detailed in Table 2 . Amplification curves resulting from 3 × 10 5 (black traces), 3 × 10 4 (red traces), 3 × 10 3 (blue traces), 3 × 10 2 (pink traces), 30 (green traces), and 0 (gray) copies of SARS-CoV-2 N gene armored RNA are depicted.

    Article Snippet: These results suggest that Taq DNA polymerase can support TaqMan RT-qPCR analyses of RNA in one-enzyme reactions.

    Techniques: Quantitative RT-PCR, Amplification

    TaqMan RT-qPCR analysis of SARS-CoV-2 viral genomic RNA and RNaseP armored RNA using Taq DNA polymerase-based one-enzyme assays. CDC SARS-CoV-2 N gene assays, N1, N2, and N3, and RNaseP assay were performed using Taq DNA polymerase from either NEB (panels A-H) or Thermo Fisher (panels I-P). Assays were performed either using the companion commercial buffer (panels A-D and panels I-L) or using Gen 6 A buffer (panels E-H and panels M-P). Amplification curves from 6000 (black traces), 600 (red traces), 60 (blue traces), 6 (pink traces), and 0 (gray traces) copies of viral genomic RNA are depicted in panels A-C, E-G, I-K, and M-O. Amplification curves from 3 × 10 5 (black traces), 3 × 10 4 (red traces), 3 × 10 3 (blue traces), 3 × 10 2 (pink traces) and 0 (gray traces) copies of armored RNaseP RNA are depicted in panes D, H, L, and P.

    Journal: bioRxiv

    Article Title: One enzyme reverse transcription qPCR using Taq DNA polymerase

    doi: 10.1101/2020.05.27.120238

    Figure Lengend Snippet: TaqMan RT-qPCR analysis of SARS-CoV-2 viral genomic RNA and RNaseP armored RNA using Taq DNA polymerase-based one-enzyme assays. CDC SARS-CoV-2 N gene assays, N1, N2, and N3, and RNaseP assay were performed using Taq DNA polymerase from either NEB (panels A-H) or Thermo Fisher (panels I-P). Assays were performed either using the companion commercial buffer (panels A-D and panels I-L) or using Gen 6 A buffer (panels E-H and panels M-P). Amplification curves from 6000 (black traces), 600 (red traces), 60 (blue traces), 6 (pink traces), and 0 (gray traces) copies of viral genomic RNA are depicted in panels A-C, E-G, I-K, and M-O. Amplification curves from 3 × 10 5 (black traces), 3 × 10 4 (red traces), 3 × 10 3 (blue traces), 3 × 10 2 (pink traces) and 0 (gray traces) copies of armored RNaseP RNA are depicted in panes D, H, L, and P.

    Article Snippet: These results suggest that Taq DNA polymerase can support TaqMan RT-qPCR analyses of RNA in one-enzyme reactions.

    Techniques: Quantitative RT-PCR, Amplification