taq dna polymerase supertaq  (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:
    1200
    Size:
    20 000 units
    Category:
    Thermostable DNA Polymerases
    Score:
    85
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    New England Biolabs taq dna polymerase supertaq
    Taq DNA Polymerase with Standard Taq Buffer
    Taq DNA Polymerase with Standard Taq Buffer 20 000 units
    https://www.bioz.com/result/taq dna polymerase supertaq/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    taq dna polymerase supertaq - by Bioz Stars, 2020-01
    99/100 stars

    Images

    1) Product Images from "A polymerase engineered for bisulfite sequencing"

    Article Title: A polymerase engineered for bisulfite sequencing

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkv798

    DNA modification and bypass during bisulfite treatment. ( A ) Scheme of the chemical reaction of cytidine: Treatment with bisulfite generates the non-aromatic, non-planar 5,6-dihydrouridine-6-sulfonate (dhU6S), which decomposes to uracil upon treatment with base (and heat). ( B ) Primer extension activity of different polymerases (Taq, 5D4, 3A10, E10, TgoT) on template T1 either unmodified (C), bisulfite-treated and desulfonated (Reagent 1, 80°C, 20 min) (converting dC to dU) (D) or bisulfite-treated (Reagent 1) but not desulphonated (converting dC to dhU6S) (S). Polymerases 5D4 and 3A10 are able to generate full-length (+20) products even from the non-desulphonated template (S). ( C ) Time-course comparison of primer extension activity of Taq and 5D4 on T1 either unmodified (C) or bisulfite-treated with (D) or without desulphonation (S). (P: primer).
    Figure Legend Snippet: DNA modification and bypass during bisulfite treatment. ( A ) Scheme of the chemical reaction of cytidine: Treatment with bisulfite generates the non-aromatic, non-planar 5,6-dihydrouridine-6-sulfonate (dhU6S), which decomposes to uracil upon treatment with base (and heat). ( B ) Primer extension activity of different polymerases (Taq, 5D4, 3A10, E10, TgoT) on template T1 either unmodified (C), bisulfite-treated and desulfonated (Reagent 1, 80°C, 20 min) (converting dC to dU) (D) or bisulfite-treated (Reagent 1) but not desulphonated (converting dC to dhU6S) (S). Polymerases 5D4 and 3A10 are able to generate full-length (+20) products even from the non-desulphonated template (S). ( C ) Time-course comparison of primer extension activity of Taq and 5D4 on T1 either unmodified (C) or bisulfite-treated with (D) or without desulphonation (S). (P: primer).

    Techniques Used: Modification, Activity Assay

    Amplification of 24 individual human genomic loci. Comparison of PCR performance of Taq (top) panel, with two different Taq/5D4 blends (5/1 (middle panel); 10/1 (bottom panel)) on the amplification of 24 different promotor regions in bisulfite-treated and fully desulphonated human genomic DNA. Both blends are able to amplify a significantly larger number of loci than Taq alone and together enable amplification of 18 out of 24 loci (75%).
    Figure Legend Snippet: Amplification of 24 individual human genomic loci. Comparison of PCR performance of Taq (top) panel, with two different Taq/5D4 blends (5/1 (middle panel); 10/1 (bottom panel)) on the amplification of 24 different promotor regions in bisulfite-treated and fully desulphonated human genomic DNA. Both blends are able to amplify a significantly larger number of loci than Taq alone and together enable amplification of 18 out of 24 loci (75%).

    Techniques Used: Amplification, Polymerase Chain Reaction

    PCR amplification of bisulfite-treated plasmid templates. PCR amplification of bisulfite-treated high GC and low GC content templates ranging from 200–600 bp in size using fully desulphonated templates and three different 5D4/Taq blends (1/10, 1/5, 1/1) with progressively lower Taq content and Taq alone on low dC content plasmid regions (top panel) and high dC content plasmid regions (bottom panel). On templates with low dC content (and hence lower levels of dU and residual dhU6S adducts post bisulfite treatment and desulphonation) either Taq or Taq/5D4 polymerase blends with a high amount of Taq perform best. In contrast on the higher dC content templates only blends containing 5D4 yield amplicons with Taq/5D4 blends (10/1; 5/1) superior to 5D4/Taq 1/1 blend, while Taq alone does not yield any amplification products. Thus only 5D4/Taq blends are able to copy the high GC content templates indicating that the blended enzymes are more efficient at copying templates containing sporadic dUs (and dhU6S adducts) and dU homopolymer stretches. Low molecular weight bands result from primer-dimer formation. (M: E-Gel ® Low Range Quantitative DNA Ladder).
    Figure Legend Snippet: PCR amplification of bisulfite-treated plasmid templates. PCR amplification of bisulfite-treated high GC and low GC content templates ranging from 200–600 bp in size using fully desulphonated templates and three different 5D4/Taq blends (1/10, 1/5, 1/1) with progressively lower Taq content and Taq alone on low dC content plasmid regions (top panel) and high dC content plasmid regions (bottom panel). On templates with low dC content (and hence lower levels of dU and residual dhU6S adducts post bisulfite treatment and desulphonation) either Taq or Taq/5D4 polymerase blends with a high amount of Taq perform best. In contrast on the higher dC content templates only blends containing 5D4 yield amplicons with Taq/5D4 blends (10/1; 5/1) superior to 5D4/Taq 1/1 blend, while Taq alone does not yield any amplification products. Thus only 5D4/Taq blends are able to copy the high GC content templates indicating that the blended enzymes are more efficient at copying templates containing sporadic dUs (and dhU6S adducts) and dU homopolymer stretches. Low molecular weight bands result from primer-dimer formation. (M: E-Gel ® Low Range Quantitative DNA Ladder).

    Techniques Used: Polymerase Chain Reaction, Amplification, Plasmid Preparation, Gas Chromatography, Molecular Weight

    Degree of methylation of individual CpG sites. Promoter regions of four genes ( A — prkcdbp , B — dab2ip , C — ptgs2 , D — ezh2) were amplified with either Taq ot 5D4/Taq blends, using bisulfide-treated genomic DNA from normal cells or LNCapP cells as a template and subjected to deep sequencing. Cyan—methylated CpGs, orange—unmethylated CpGs. Individual CpGs are numbered starting from the 5′ end of the amplicon.
    Figure Legend Snippet: Degree of methylation of individual CpG sites. Promoter regions of four genes ( A — prkcdbp , B — dab2ip , C — ptgs2 , D — ezh2) were amplified with either Taq ot 5D4/Taq blends, using bisulfide-treated genomic DNA from normal cells or LNCapP cells as a template and subjected to deep sequencing. Cyan—methylated CpGs, orange—unmethylated CpGs. Individual CpGs are numbered starting from the 5′ end of the amplicon.

    Techniques Used: Methylation, Amplification, Sequencing

    Related Articles

    Clone Assay:

    Article Title: Growth of Dehalobacter and Dehalococcoides spp. during Degradation of Chlorinated Ethanes
    Article Snippet: Paragraph title: 16S rRNA gene cloning and PCR-DGGE. ... PCR was performed in triplicate 50-μl reactions containing 1× ThermoPol PCR buffer (New England Biolabs, Mississauga, Ontario, Canada), 0.5 μM concentrations of each primer, 0.5 mM deoxynucleoside triphosphates, 1.5 U of Taq DNA polymerase (New Englands Biolabs), and 50 ng of DNA.

    GWAS:

    Article Title: Identification of Quantitative Trait Loci and Candidate Genes for Maize Starch Granule Size through Association Mapping
    Article Snippet: Three candidate genes, GRMZM2G419655 , GRMZM2G419660 , and GRMZM2G511067 , were selected for sequencing based on the GWAS. .. PCR reaction mixes (20 µl) contained 1 µl of NEB (New England BioLabs Inc., Ipswich, MA, USA) Taq DNA Polymerase, 4 µl of 5× NEB PCR Buffer, 0.5 µl of dNTP mixture, 0.5 µl each of the two primers, and 1 µl of template DNA.

    Amplification:

    Article Title: Direct Selection of RNA Beacon Aptamers
    Article Snippet: 5 nmol (~3 x 1015 molecules) of DM001 were amplified in a total volume of 40 mL that was divided into two 96 well plates. .. Reactions contained 0.125 μM DM001, 1 μM DM003, 1 μM DM004 , 200 μM dNTPs, 1X buffer (provided with enzyme) and 12.5 U/mL Taq DNA polymerase (New England Biolabs).

    Article Title: Molecular Analysis of the Enterobacter sakazakii O-Antigen Gene Locus
    Article Snippet: These genes were chosen based on sequence analysis that revealed that these targets were highly conserved. .. PCR amplification was performed with 50-μl volumes containing 1× PCR buffer containing 1.5 mM MgCl2 (New England Biolabs, Herts, United Kingdom), 100 mM deoxynucleoside triphosphates, 0.1 μM of each primer (MWG Biotech AG, Ebersberg, Germany), 2.5 U Taq DNA polymerase (New England Biolabs, Herts, United Kingdom), 100 ng template DNA, and PCR-grade water (Invitrogen, CA). .. Thermal PCR conditions for O:1 were 95°C for 1 min, followed by 35 cycles of 94°C for 1 min, 56°C for 1 min (annealing), and 72°C for 1 min with a final extension of 72°C for 5 min, while O:2 PCR was annealed at 51°C for 1 min. Amplified products were analyzed by electrophoresis through a 1% (wt/vol) agarose gel at 80 V for 60 min in 1× TBE.

    Article Title: Growth of Dehalobacter and Dehalococcoides spp. during Degradation of Chlorinated Ethanes
    Article Snippet: Bacterial 16S rRNA genes were selectively amplified from the purified DNA by PCR using the forward primer 27f and the reverse primer 1492R ( ). .. PCR was performed in triplicate 50-μl reactions containing 1× ThermoPol PCR buffer (New England Biolabs, Mississauga, Ontario, Canada), 0.5 μM concentrations of each primer, 0.5 mM deoxynucleoside triphosphates, 1.5 U of Taq DNA polymerase (New Englands Biolabs), and 50 ng of DNA.

    Article Title: Genetic variations among three major ethnic groups in Nigeria using RAPD
    Article Snippet: Agarose from Cleaver Scientific Limited, Quickload 100bp DNA ladder from New England Biolabs Inc., PCR Master-mix with standard buffer (Quick-Ld2X) containing Taq polymerase was also from New England Biolabs Inc. and the oligonucleotide primers were from Inqaba, South Africa. .. Five millimeters of blood were obtained from each donor, placed in tubes containing anti-coagulant (EDTA) and stored at -20°C prior to DNA extraction.

    Article Title: Automated selection of aptamers against cholangiocarcinoma cells on an integrated microfluidic platform
    Article Snippet: The gauge pressure and frequency were then regulated such that an aliquot of the supernatant was transferred to the PCR chamber. .. The PCR chamber was preloaded with 0.5 μ l of the forward primer (5′-GGCAGGAAGACAAACA-3′, 0.5 μ M), 0.5 μ l of the reverse primer (5′-ACAGCACCACAGACCA-3′, 0.5 μ M), 0.5 μ l of deoxynucleotide triphosphates (0.2 mM), 2 μ l of ssDNA (0.1 μ M) or the previous amplified product from the last round of Cell-SELEX, 0.125 μ l of Taq DNA polymerase (New England Biolabs, USA), and double-distilled water (ddH2 O) to a final volume of 25 μ l. The PCR was initiated at 95 °C for 10 min followed by 30 cycles of 95 °C for 30 s, 63 °C for 15 s, and 72 °C for 30 s. A final extension step at 72 °C for 10 min was also incorporated into the protocol. .. The amplified products were then transferred to the positive selection chamber, and the gauge pressure and frequency were fine-tuned such that an aliquot of the amplified product was used for the next round of SELEX, with the remainder collected via the waste collection chamber.

    Article Title: Promoter Activation by CII, a Potent Transcriptional Activator from Bacteriophage 186
    Article Snippet: CII145 was amplified from pZS45-CII145 by high fidelity PCR using primers 289 and RSP (see for sequences). .. The resulting PCR product was used as template for error-prone PCR with Taq DNA polymerase (New England Biolabs) and MgCl2 supplemented to a total concentration of 6 m m .

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Nucleotide analogue mutagenesis was carried out in the presence of 2 and 20 μM 8-oxo-2′-deoxyguanosine (8-oxo-dGTP) and 6-(2-deoxy-β- d -ribofuranosyl)-3,4-dihydro-8H-pyrimido-(4,5-c)(1,2)oxazin-7-one (dPTP) ( ). .. Using the plasmid pZE21:: tyrA WT as template, 10, 20, and 30 amplification cycles with the primers mentioned above were performed using Taq DNA polymerase (New England Biolabs). .. The 1.1-kbp PCR products were gel purified, and the mutated tyrA genes were amplified in a second PCR under regular conditions.

    Article Title: Post-crystallization improvement of RNA crystal diffraction quality
    Article Snippet: Oligonucleotides for PCR amplification. .. Taq DNA polymerase, 5,000 U/mL (New England Biolabs).

    Article Title: Genetic variants in the enhancer region of the thymidylate synthase gene in the Chilean population
    Article Snippet: Two oligonucleotide primers were synthesized specifically to amplify the polymorphic region of TSER (sense primer 5′-GTGGCTCCTGCGT TTCCCCC-3′ and antisense primer 5′-GCTCCGAGC CGG CCACAGGCATGGCGCGG-3′) [ ]. .. Amplification reactions were performed in a 25-µl volume containing 0.2 µg genomic DNA, 0.2 m m dNTPs , 0.5 µ m of each primer, 1.0 units of Taq polymerase (New England BioLabs, Ipswich, MA, USA) and 2.5 µl of 10 × Taq PCR buffer. .. Cycling conditions were: one cycle at 94 °C for 5 min with hot start; 30 cycles of 40 s at 94 °C, 1 min at 62 °C and 40 s at 72 °C; and one elongation cycle for 5 min at 72 °C.

    Article Title: Targeted gene inactivation in zebrafish using engineered zinc finger nucleases
    Article Snippet: Colonies from the desired selection plates were washed off the plates and their plasmid DNA was recovered as a pool as previously described . .. Each pool of individual fingers was amplified from the plasmid DNA by PCR: 50 μl reaction with 1 units NEB DNA Taq polymerase using 50 ng plasmid DNA template and 1 mM each primer ( ); denature for 1 min at 95°C, 28 cycles consisting of 94°C for 20 sec, 55°C for 20 sec and 72°C for 20 sec, followed by a final extension at 72°C for 6 min. .. The PCR products were run on a 1.5% agarose TAE gel, and the appropriate bands were excised and purified using a Qiagen PCR purification kit.

    Article Title: Pseudomonas aeruginosa AlgR Phosphorylation Modulates Rhamnolipid Production and Motility
    Article Snippet: Electrophoretic mobility shift assays (EMSAs) were performed using purified AlgR proteins, as described previously ( ). .. DNA fragments were prepared by PCR amplification of PAO1 chromosomal DNA with Taq polymerase (New England BioLabs) and the oligonucleotides listed in for fimU and rhlA . .. Oligonucleotides were 5′-end labeled with [γ-32 P]ATP (6,000 Ci/mmol) (Perkin-Elmer) with polynucleotide kinase (New England BioLabs).

    Article Title: Evidence for Acquisition of Legionella Type IV Secretion Substrates via Interdomain Horizontal Gene Transfer
    Article Snippet: Finally, we screened the individual plate by the same PCR technique to identify the mutant of interest. .. PCRs were performed using Taq polymerase (New England Biolabs, Massachusetts) with the following protocol: denaturation at 95°C for 5 min, followed by 35 cycles of denaturation at 95°C for 10 s, annealing at 68°C for 30 s, and amplification at 72°C for 3 min. .. The PCR products were then sequenced to confirm that the transposon insertion was in fact located in the gene of interest.

    Synthesized:

    Article Title: Genetic variants in the enhancer region of the thymidylate synthase gene in the Chilean population
    Article Snippet: Two oligonucleotide primers were synthesized specifically to amplify the polymorphic region of TSER (sense primer 5′-GTGGCTCCTGCGT TTCCCCC-3′ and antisense primer 5′-GCTCCGAGC CGG CCACAGGCATGGCGCGG-3′) [ ]. .. Amplification reactions were performed in a 25-µl volume containing 0.2 µg genomic DNA, 0.2 m m dNTPs , 0.5 µ m of each primer, 1.0 units of Taq polymerase (New England BioLabs, Ipswich, MA, USA) and 2.5 µl of 10 × Taq PCR buffer.

    TA Cloning:

    Article Title: Growth of Dehalobacter and Dehalococcoides spp. during Degradation of Chlorinated Ethanes
    Article Snippet: PCR was performed in triplicate 50-μl reactions containing 1× ThermoPol PCR buffer (New England Biolabs, Mississauga, Ontario, Canada), 0.5 μM concentrations of each primer, 0.5 mM deoxynucleoside triphosphates, 1.5 U of Taq DNA polymerase (New Englands Biolabs), and 50 ng of DNA. .. The conditions used for PCR amplification were as follows: initial denaturation at 94°C for 5 min and then 25 cycles of (denaturation 94°C for 30 s, primer annealing at 52°C for 30 s, and chain extension for 1 min at 72°C), followed by a final extension at 72°C for 10 min. A PTC-200 thermocycler (MJ Research, Inc., Waltham, MA) was used for PCR.

    Real-time Polymerase Chain Reaction:

    Article Title: Field evaluation of a real time loop-mediated isothermal amplification assay (RealAmp) for malaria diagnosis in Cruzeiro do Sul, Acre, Brazil
    Article Snippet: Nested PCR was conducted in cases of disagreement between study microscopy and the real-time PCR results. .. Reactions were performed in 20 μL total volume containing 1X buffer, 2.5 mM MgCl2 , 200 mM dNTPs, 200 nM primers, and 1.25 units of Taq Polymerase (New England Biolabs, Ipswich, MA).

    Incubation:

    Article Title: Promoter Activation by CII, a Potent Transcriptional Activator from Bacteriophage 186
    Article Snippet: The resulting PCR product was used as template for error-prone PCR with Taq DNA polymerase (New England Biolabs) and MgCl2 supplemented to a total concentration of 6 m m . .. The product of error-prone PCR was subcloned back into pZS45-CII145 via BamHI and XhoI restriction sites and transformed into screening strain IM18 by electroporation.

    Transformation Assay:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Using the plasmid pZE21:: tyrA WT as template, 10, 20, and 30 amplification cycles with the primers mentioned above were performed using Taq DNA polymerase (New England Biolabs). .. The 1.1-kbp PCR products were gel purified, and the mutated tyrA genes were amplified in a second PCR under regular conditions.

    Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
    Article Snippet: E. coli transformation, plasmid isolation, and recombinant techniques were performed using standard methods ( ). .. Restriction enzymes, T4 DNA ligase, and Taq DNA polymerase were purchased from NEB.

    Article Title: The Alternative Sigma Factor ?H Is Required for Toxin Gene Expression by Bacillus anthracis
    Article Snippet: Preparation of plasmid DNA from E. coli , transformation of E. coli , and recombinant DNA techniques were performed using standard procedures ( ). .. Restriction enzymes and T4 DNA ligase were purchased from Promega (Madison, WI) and Fisher Scientific, and Taq DNA polymerase was purchased from New England Biolabs (Beverly, MA).

    RNA Binding Assay:

    Article Title: Post-crystallization improvement of RNA crystal diffraction quality
    Article Snippet: Taq DNA polymerase, 5,000 U/mL (New England Biolabs). .. Taq DNA polymerase, 5,000 U/mL (New England Biolabs).

    Countercurrent Chromatography:

    Article Title: Eco-Evolutionary Processes Generating Diversity Among Bottlenose Dolphin, Tursiops truncatus, Populations off Baja California, Mexico
    Article Snippet: Fragments of 480 bp from the mtDNA control region, tRNA proline end, were analysed for 250 samples (see Table ). .. The PCRs were performed in 25µL volumes consisting of 10 mM Tris–HCl, 50 mM KCl, 2.5 mM MgCl2 , 0.25 mM each dNTP, 0.12 µM each primer: L15812 (TRO): 5′ CCT CCC TAA GAC TCA AGG AAG 3′ and (D): 5′ CCT GAA GTA AGA ACC AGA TG 3′ (Rosel et al. ), 1.25 U of Taq DNA polymerase (NEB, UK), and approximately 50 ng of genomic DNA. .. The cycle profile was 5 min at 95 °C, followed by 36 amplification cycles of 45 s at 48 °C, 1 min at 72 °C and 45 s at 94 °C and a final elongation step of 10 min at 72 °C.

    Electroporation:

    Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
    Article Snippet: Non-methylated plasmid DNA for electroporation into B. anthracis ( ; ) was obtained from E. coli GM2163 cells. .. Restriction enzymes, T4 DNA ligase, and Taq DNA polymerase were purchased from NEB.

    Concentration Assay:

    Article Title: Promoter Activation by CII, a Potent Transcriptional Activator from Bacteriophage 186
    Article Snippet: CII145 was amplified from pZS45-CII145 by high fidelity PCR using primers 289 and RSP (see for sequences). .. The resulting PCR product was used as template for error-prone PCR with Taq DNA polymerase (New England Biolabs) and MgCl2 supplemented to a total concentration of 6 m m . .. The product of error-prone PCR was subcloned back into pZS45-CII145 via BamHI and XhoI restriction sites and transformed into screening strain IM18 by electroporation.

    Buffer Exchange:

    Article Title: Discovery of Bacterial sRNAs by High-Throughput Sequencing
    Article Snippet: Taq DNA Polymerase (with 10× Buffer) (NEB, Ipswich, MA). .. Taq DNA Polymerase (with 10× Buffer) (NEB, Ipswich, MA).

    Northern Blot:

    Article Title: Genetic variants in the enhancer region of the thymidylate synthase gene in the Chilean population
    Article Snippet: The SJH and CLC are located in the northern and eastern parts of Santiago, respectively. .. Amplification reactions were performed in a 25-µl volume containing 0.2 µg genomic DNA, 0.2 m m dNTPs , 0.5 µ m of each primer, 1.0 units of Taq polymerase (New England BioLabs, Ipswich, MA, USA) and 2.5 µl of 10 × Taq PCR buffer.

    Hemagglutination Assay:

    Article Title: Mechanisms of pseudosubstrate inhibition of the anaphase promoting complex by Acm1
    Article Snippet: The sense primer annealed to the GAL4 DNA-binding domain and HA tag and extended to the CDH1 start codon. .. EP-PCR conditions were as follows: 2 μg/ml CDH1- Δ 200-URA3 -pAS2, 0.2 μM of each primer, 1 mM dTTP, 1 mM dCTP, 0.2 mM dATP, 0.2 mM dGTP, 6 mM MgCl2 , 50 μM MnCl2 , 1 × PCR buffer (−MgCl2 ) (Invitrogen) and 0.5 U/μl Taq polymerase (New England Biolabs).

    DNA Sequencing:

    Article Title: Growth of Dehalobacter and Dehalococcoides spp. during Degradation of Chlorinated Ethanes
    Article Snippet: PCR was performed in triplicate 50-μl reactions containing 1× ThermoPol PCR buffer (New England Biolabs, Mississauga, Ontario, Canada), 0.5 μM concentrations of each primer, 0.5 mM deoxynucleoside triphosphates, 1.5 U of Taq DNA polymerase (New Englands Biolabs), and 50 ng of DNA. .. Triplicate reactions were combined and cloned with the TOPO TA cloning kit (Invitrogen Corp., Carlsbad, CA) according to the manufacturer's protocol.

    Sequencing:

    Article Title: Discovery of Bacterial sRNAs by High-Throughput Sequencing
    Article Snippet: Taq DNA Polymerase (with 10× Buffer) (NEB, Ipswich, MA). .. Taq DNA Polymerase (with 10× Buffer) (NEB, Ipswich, MA).

    Article Title: Molecular Analysis of the Enterobacter sakazakii O-Antigen Gene Locus
    Article Snippet: PCR amplification was performed with 50-μl volumes containing 1× PCR buffer containing 1.5 mM MgCl2 (New England Biolabs, Herts, United Kingdom), 100 mM deoxynucleoside triphosphates, 0.1 μM of each primer (MWG Biotech AG, Ebersberg, Germany), 2.5 U Taq DNA polymerase (New England Biolabs, Herts, United Kingdom), 100 ng template DNA, and PCR-grade water (Invitrogen, CA). .. PCR amplification was performed with 50-μl volumes containing 1× PCR buffer containing 1.5 mM MgCl2 (New England Biolabs, Herts, United Kingdom), 100 mM deoxynucleoside triphosphates, 0.1 μM of each primer (MWG Biotech AG, Ebersberg, Germany), 2.5 U Taq DNA polymerase (New England Biolabs, Herts, United Kingdom), 100 ng template DNA, and PCR-grade water (Invitrogen, CA).

    Article Title: Growth of Dehalobacter and Dehalococcoides spp. during Degradation of Chlorinated Ethanes
    Article Snippet: PCR was performed in triplicate 50-μl reactions containing 1× ThermoPol PCR buffer (New England Biolabs, Mississauga, Ontario, Canada), 0.5 μM concentrations of each primer, 0.5 mM deoxynucleoside triphosphates, 1.5 U of Taq DNA polymerase (New Englands Biolabs), and 50 ng of DNA. .. Triplicate reactions were combined and cloned with the TOPO TA cloning kit (Invitrogen Corp., Carlsbad, CA) according to the manufacturer's protocol.

    Article Title: Identification of Quantitative Trait Loci and Candidate Genes for Maize Starch Granule Size through Association Mapping
    Article Snippet: Paragraph title: Sequencing and candidate gene association analysis ... PCR reaction mixes (20 µl) contained 1 µl of NEB (New England BioLabs Inc., Ipswich, MA, USA) Taq DNA Polymerase, 4 µl of 5× NEB PCR Buffer, 0.5 µl of dNTP mixture, 0.5 µl each of the two primers, and 1 µl of template DNA.

    Binding Assay:

    Article Title: Automated selection of aptamers against cholangiocarcinoma cells on an integrated microfluidic platform
    Article Snippet: With the aim of reducing the non-specific binding of the aptamers, one additional round of negative selection with MMNK-1 cell lines was performed with the 6th SELEX round PCR product for the SNU-478 and HuCCT-1 cell lines. .. The PCR chamber was preloaded with 0.5 μ l of the forward primer (5′-GGCAGGAAGACAAACA-3′, 0.5 μ M), 0.5 μ l of the reverse primer (5′-ACAGCACCACAGACCA-3′, 0.5 μ M), 0.5 μ l of deoxynucleotide triphosphates (0.2 mM), 2 μ l of ssDNA (0.1 μ M) or the previous amplified product from the last round of Cell-SELEX, 0.125 μ l of Taq DNA polymerase (New England Biolabs, USA), and double-distilled water (ddH2 O) to a final volume of 25 μ l. The PCR was initiated at 95 °C for 10 min followed by 30 cycles of 95 °C for 30 s, 63 °C for 15 s, and 72 °C for 30 s. A final extension step at 72 °C for 10 min was also incorporated into the protocol.

    Article Title: Pseudomonas aeruginosa AlgR Phosphorylation Modulates Rhamnolipid Production and Motility
    Article Snippet: DNA fragments were prepared by PCR amplification of PAO1 chromosomal DNA with Taq polymerase (New England BioLabs) and the oligonucleotides listed in for fimU and rhlA . .. The templates for the rhlAΔ1 (primers rhlA SDM delta RB1 F and rhlA SDM WT RB2R), rhlAΔ2 (primers rhlA SDM WT RB1 F and rhlA SDM delta RB2R), and rhlAΔ1Δ2 (primers rhlA SDM delta RB1 F and rhlA SDM delta RB2R) oligonucleotides were created using site-directed mutagenesis of the pGEMT- rhlA prom vector.

    DNA Extraction:

    Article Title: Genetic variations among three major ethnic groups in Nigeria using RAPD
    Article Snippet: Materials: Genomic DNA isolation kits were purchased from Norgen Biotek Corporation, (Thorold, Canada). .. Agarose from Cleaver Scientific Limited, Quickload 100bp DNA ladder from New England Biolabs Inc., PCR Master-mix with standard buffer (Quick-Ld2X) containing Taq polymerase was also from New England Biolabs Inc. and the oligonucleotide primers were from Inqaba, South Africa.

    Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
    Article Snippet: Paragraph title: DNA Isolation and Manipulation ... Restriction enzymes, T4 DNA ligase, and Taq DNA polymerase were purchased from NEB.

    Article Title: The Alternative Sigma Factor ?H Is Required for Toxin Gene Expression by Bacillus anthracis
    Article Snippet: Paragraph title: DNA isolation and manipulation. ... Restriction enzymes and T4 DNA ligase were purchased from Promega (Madison, WI) and Fisher Scientific, and Taq DNA polymerase was purchased from New England Biolabs (Beverly, MA).

    Mutagenesis:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Nucleotide analogue mutagenesis was carried out in the presence of 2 and 20 μM 8-oxo-2′-deoxyguanosine (8-oxo-dGTP) and 6-(2-deoxy-β- d -ribofuranosyl)-3,4-dihydro-8H-pyrimido-(4,5-c)(1,2)oxazin-7-one (dPTP) ( ). .. Using the plasmid pZE21:: tyrA WT as template, 10, 20, and 30 amplification cycles with the primers mentioned above were performed using Taq DNA polymerase (New England Biolabs).

    Article Title: Mechanisms of pseudosubstrate inhibition of the anaphase promoting complex by Acm1
    Article Snippet: Paragraph title: EP-PCR mutagenesis of CDH1- Δ 200 and yeast two-hybrid screening ... EP-PCR conditions were as follows: 2 μg/ml CDH1- Δ 200-URA3 -pAS2, 0.2 μM of each primer, 1 mM dTTP, 1 mM dCTP, 0.2 mM dATP, 0.2 mM dGTP, 6 mM MgCl2 , 50 μM MnCl2 , 1 × PCR buffer (−MgCl2 ) (Invitrogen) and 0.5 U/μl Taq polymerase (New England Biolabs).

    Article Title: Pseudomonas aeruginosa AlgR Phosphorylation Modulates Rhamnolipid Production and Motility
    Article Snippet: DNA fragments were prepared by PCR amplification of PAO1 chromosomal DNA with Taq polymerase (New England BioLabs) and the oligonucleotides listed in for fimU and rhlA . .. DNA fragments were prepared by PCR amplification of PAO1 chromosomal DNA with Taq polymerase (New England BioLabs) and the oligonucleotides listed in for fimU and rhlA .

    Article Title: Evidence for Acquisition of Legionella Type IV Secretion Substrates via Interdomain Horizontal Gene Transfer
    Article Snippet: Finally, we screened the individual plate by the same PCR technique to identify the mutant of interest. .. PCRs were performed using Taq polymerase (New England Biolabs, Massachusetts) with the following protocol: denaturation at 95°C for 5 min, followed by 35 cycles of denaturation at 95°C for 10 s, annealing at 68°C for 30 s, and amplification at 72°C for 3 min.

    Isolation:

    Article Title: Automated selection of aptamers against cholangiocarcinoma cells on an integrated microfluidic platform
    Article Snippet: The product of this final round was then used in a negative selection step with White blood cells (WBCs) isolated from whole blood (described in more detail below). .. The PCR chamber was preloaded with 0.5 μ l of the forward primer (5′-GGCAGGAAGACAAACA-3′, 0.5 μ M), 0.5 μ l of the reverse primer (5′-ACAGCACCACAGACCA-3′, 0.5 μ M), 0.5 μ l of deoxynucleotide triphosphates (0.2 mM), 2 μ l of ssDNA (0.1 μ M) or the previous amplified product from the last round of Cell-SELEX, 0.125 μ l of Taq DNA polymerase (New England Biolabs, USA), and double-distilled water (ddH2 O) to a final volume of 25 μ l. The PCR was initiated at 95 °C for 10 min followed by 30 cycles of 95 °C for 30 s, 63 °C for 15 s, and 72 °C for 30 s. A final extension step at 72 °C for 10 min was also incorporated into the protocol.

    Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
    Article Snippet: E. coli transformation, plasmid isolation, and recombinant techniques were performed using standard methods ( ). .. Restriction enzymes, T4 DNA ligase, and Taq DNA polymerase were purchased from NEB.

    Article Title: The Alternative Sigma Factor ?H Is Required for Toxin Gene Expression by Bacillus anthracis
    Article Snippet: Extraction of chromosomal DNA from B. anthracis cultures was carried out using a Mo Bio genomic isolation kit (Mo Bio Laboratories, Solana Beach, CA). .. Restriction enzymes and T4 DNA ligase were purchased from Promega (Madison, WI) and Fisher Scientific, and Taq DNA polymerase was purchased from New England Biolabs (Beverly, MA).

    Article Title: Genetic variants in the enhancer region of the thymidylate synthase gene in the Chilean population
    Article Snippet: To determine the TSER variants, DNA was isolated by routine methods and amplified by polymerase chain reaction (PCR) [ ]. .. Amplification reactions were performed in a 25-µl volume containing 0.2 µg genomic DNA, 0.2 m m dNTPs , 0.5 µ m of each primer, 1.0 units of Taq polymerase (New England BioLabs, Ipswich, MA, USA) and 2.5 µl of 10 × Taq PCR buffer.

    Size-exclusion Chromatography:

    Article Title: Targeted gene inactivation in zebrafish using engineered zinc finger nucleases
    Article Snippet: Colonies from the desired selection plates were washed off the plates and their plasmid DNA was recovered as a pool as previously described . .. Each pool of individual fingers was amplified from the plasmid DNA by PCR: 50 μl reaction with 1 units NEB DNA Taq polymerase using 50 ng plasmid DNA template and 1 mM each primer ( ); denature for 1 min at 95°C, 28 cycles consisting of 94°C for 20 sec, 55°C for 20 sec and 72°C for 20 sec, followed by a final extension at 72°C for 6 min. .. The PCR products were run on a 1.5% agarose TAE gel, and the appropriate bands were excised and purified using a Qiagen PCR purification kit.

    Microscopy:

    Article Title: Field evaluation of a real time loop-mediated isothermal amplification assay (RealAmp) for malaria diagnosis in Cruzeiro do Sul, Acre, Brazil
    Article Snippet: Nested PCR was conducted in cases of disagreement between study microscopy and the real-time PCR results. .. Reactions were performed in 20 μL total volume containing 1X buffer, 2.5 mM MgCl2 , 200 mM dNTPs, 200 nM primers, and 1.25 units of Taq Polymerase (New England Biolabs, Ipswich, MA).

    Purification:

    Article Title: Growth of Dehalobacter and Dehalococcoides spp. during Degradation of Chlorinated Ethanes
    Article Snippet: Bacterial 16S rRNA genes were selectively amplified from the purified DNA by PCR using the forward primer 27f and the reverse primer 1492R ( ). .. PCR was performed in triplicate 50-μl reactions containing 1× ThermoPol PCR buffer (New England Biolabs, Mississauga, Ontario, Canada), 0.5 μM concentrations of each primer, 0.5 mM deoxynucleoside triphosphates, 1.5 U of Taq DNA polymerase (New Englands Biolabs), and 50 ng of DNA.

    Article Title: Eco-Evolutionary Processes Generating Diversity Among Bottlenose Dolphin, Tursiops truncatus, Populations off Baja California, Mexico
    Article Snippet: The PCRs were performed in 25µL volumes consisting of 10 mM Tris–HCl, 50 mM KCl, 2.5 mM MgCl2 , 0.25 mM each dNTP, 0.12 µM each primer: L15812 (TRO): 5′ CCT CCC TAA GAC TCA AGG AAG 3′ and (D): 5′ CCT GAA GTA AGA ACC AGA TG 3′ (Rosel et al. ), 1.25 U of Taq DNA polymerase (NEB, UK), and approximately 50 ng of genomic DNA. .. The PCRs were performed in 25µL volumes consisting of 10 mM Tris–HCl, 50 mM KCl, 2.5 mM MgCl2 , 0.25 mM each dNTP, 0.12 µM each primer: L15812 (TRO): 5′ CCT CCC TAA GAC TCA AGG AAG 3′ and (D): 5′ CCT GAA GTA AGA ACC AGA TG 3′ (Rosel et al. ), 1.25 U of Taq DNA polymerase (NEB, UK), and approximately 50 ng of genomic DNA.

    Article Title: Mechanisms of pseudosubstrate inhibition of the anaphase promoting complex by Acm1
    Article Snippet: EP-PCR conditions were as follows: 2 μg/ml CDH1- Δ 200-URA3 -pAS2, 0.2 μM of each primer, 1 mM dTTP, 1 mM dCTP, 0.2 mM dATP, 0.2 mM dGTP, 6 mM MgCl2 , 50 μM MnCl2 , 1 × PCR buffer (−MgCl2 ) (Invitrogen) and 0.5 U/μl Taq polymerase (New England Biolabs). .. After 16 cycles of amplification, 1 ml of product was purified using a Qiagen PCR purification column (Qiagen, Inc.) and co-transformed into YJB1212 with ∼2.5 μg of gapped CDH1- Δ 200-URA3 -pAS2 vector.

    Article Title: Pseudomonas aeruginosa AlgR Phosphorylation Modulates Rhamnolipid Production and Motility
    Article Snippet: Electrophoretic mobility shift assays (EMSAs) were performed using purified AlgR proteins, as described previously ( ). .. DNA fragments were prepared by PCR amplification of PAO1 chromosomal DNA with Taq polymerase (New England BioLabs) and the oligonucleotides listed in for fimU and rhlA .

    Polymerase Chain Reaction:

    Article Title: Discovery of Bacterial sRNAs by High-Throughput Sequencing
    Article Snippet: Paragraph title: 2.6. PCR ... Taq DNA Polymerase (with 10× Buffer) (NEB, Ipswich, MA).

    Article Title: Direct Selection of RNA Beacon Aptamers
    Article Snippet: Reactions contained 0.125 μM DM001, 1 μM DM003, 1 μM DM004 , 200 μM dNTPs, 1X buffer (provided with enzyme) and 12.5 U/mL Taq DNA polymerase (New England Biolabs). .. Reactions contained 0.125 μM DM001, 1 μM DM003, 1 μM DM004 , 200 μM dNTPs, 1X buffer (provided with enzyme) and 12.5 U/mL Taq DNA polymerase (New England Biolabs).

    Article Title: Molecular Analysis of the Enterobacter sakazakii O-Antigen Gene Locus
    Article Snippet: These genes were chosen based on sequence analysis that revealed that these targets were highly conserved. .. PCR amplification was performed with 50-μl volumes containing 1× PCR buffer containing 1.5 mM MgCl2 (New England Biolabs, Herts, United Kingdom), 100 mM deoxynucleoside triphosphates, 0.1 μM of each primer (MWG Biotech AG, Ebersberg, Germany), 2.5 U Taq DNA polymerase (New England Biolabs, Herts, United Kingdom), 100 ng template DNA, and PCR-grade water (Invitrogen, CA). .. Thermal PCR conditions for O:1 were 95°C for 1 min, followed by 35 cycles of 94°C for 1 min, 56°C for 1 min (annealing), and 72°C for 1 min with a final extension of 72°C for 5 min, while O:2 PCR was annealed at 51°C for 1 min. Amplified products were analyzed by electrophoresis through a 1% (wt/vol) agarose gel at 80 V for 60 min in 1× TBE.

    Article Title: Growth of Dehalobacter and Dehalococcoides spp. during Degradation of Chlorinated Ethanes
    Article Snippet: Bacterial 16S rRNA genes were selectively amplified from the purified DNA by PCR using the forward primer 27f and the reverse primer 1492R ( ). .. PCR was performed in triplicate 50-μl reactions containing 1× ThermoPol PCR buffer (New England Biolabs, Mississauga, Ontario, Canada), 0.5 μM concentrations of each primer, 0.5 mM deoxynucleoside triphosphates, 1.5 U of Taq DNA polymerase (New Englands Biolabs), and 50 ng of DNA. .. The conditions used for PCR amplification were as follows: initial denaturation at 94°C for 5 min and then 25 cycles of (denaturation 94°C for 30 s, primer annealing at 52°C for 30 s, and chain extension for 1 min at 72°C), followed by a final extension at 72°C for 10 min. A PTC-200 thermocycler (MJ Research, Inc., Waltham, MA) was used for PCR.

    Article Title: Genetic variations among three major ethnic groups in Nigeria using RAPD
    Article Snippet: Materials: Genomic DNA isolation kits were purchased from Norgen Biotek Corporation, (Thorold, Canada). .. Agarose from Cleaver Scientific Limited, Quickload 100bp DNA ladder from New England Biolabs Inc., PCR Master-mix with standard buffer (Quick-Ld2X) containing Taq polymerase was also from New England Biolabs Inc. and the oligonucleotide primers were from Inqaba, South Africa. .. All other chemicals were of molecular biology grade.

    Article Title: Automated selection of aptamers against cholangiocarcinoma cells on an integrated microfluidic platform
    Article Snippet: The gauge pressure and frequency were then regulated such that an aliquot of the supernatant was transferred to the PCR chamber. .. The PCR chamber was preloaded with 0.5 μ l of the forward primer (5′-GGCAGGAAGACAAACA-3′, 0.5 μ M), 0.5 μ l of the reverse primer (5′-ACAGCACCACAGACCA-3′, 0.5 μ M), 0.5 μ l of deoxynucleotide triphosphates (0.2 mM), 2 μ l of ssDNA (0.1 μ M) or the previous amplified product from the last round of Cell-SELEX, 0.125 μ l of Taq DNA polymerase (New England Biolabs, USA), and double-distilled water (ddH2 O) to a final volume of 25 μ l. The PCR was initiated at 95 °C for 10 min followed by 30 cycles of 95 °C for 30 s, 63 °C for 15 s, and 72 °C for 30 s. A final extension step at 72 °C for 10 min was also incorporated into the protocol. .. The amplified products were then transferred to the positive selection chamber, and the gauge pressure and frequency were fine-tuned such that an aliquot of the amplified product was used for the next round of SELEX, with the remainder collected via the waste collection chamber.

    Article Title: Promoter Activation by CII, a Potent Transcriptional Activator from Bacteriophage 186
    Article Snippet: CII145 was amplified from pZS45-CII145 by high fidelity PCR using primers 289 and RSP (see for sequences). .. The resulting PCR product was used as template for error-prone PCR with Taq DNA polymerase (New England Biolabs) and MgCl2 supplemented to a total concentration of 6 m m . .. The product of error-prone PCR was subcloned back into pZS45-CII145 via BamHI and XhoI restriction sites and transformed into screening strain IM18 by electroporation.

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Paragraph title: Error-prone PCR and selection of feedback-inhibition-resistant tyrA mutants. ... Using the plasmid pZE21:: tyrA WT as template, 10, 20, and 30 amplification cycles with the primers mentioned above were performed using Taq DNA polymerase (New England Biolabs).

    Article Title: Post-crystallization improvement of RNA crystal diffraction quality
    Article Snippet: Oligonucleotides for PCR amplification. .. Taq DNA polymerase, 5,000 U/mL (New England Biolabs).

    Article Title: Specific Roles of the iroBCDEN Genes in Virulence of an Avian Pathogenic Escherichia coli O78 Strain and in Production of Salmochelins
    Article Snippet: Two hundred ninety-eight APEC and 32 commensal fecal isolates were screened by PCR for the presence of the iroB and iroN genes. .. The PCRs were achieved using Taq DNA polymerase (New England Biolabs), at an annealing temperature of 58°C and an extension time of 1 min at 72°C for 25 cycles.

    Article Title: Eco-Evolutionary Processes Generating Diversity Among Bottlenose Dolphin, Tursiops truncatus, Populations off Baja California, Mexico
    Article Snippet: The PCRs were performed in 25µL volumes consisting of 10 mM Tris–HCl, 50 mM KCl, 2.5 mM MgCl2 , 0.25 mM each dNTP, 0.12 µM each primer: L15812 (TRO): 5′ CCT CCC TAA GAC TCA AGG AAG 3′ and (D): 5′ CCT GAA GTA AGA ACC AGA TG 3′ (Rosel et al. ), 1.25 U of Taq DNA polymerase (NEB, UK), and approximately 50 ng of genomic DNA. .. The PCRs were performed in 25µL volumes consisting of 10 mM Tris–HCl, 50 mM KCl, 2.5 mM MgCl2 , 0.25 mM each dNTP, 0.12 µM each primer: L15812 (TRO): 5′ CCT CCC TAA GAC TCA AGG AAG 3′ and (D): 5′ CCT GAA GTA AGA ACC AGA TG 3′ (Rosel et al. ), 1.25 U of Taq DNA polymerase (NEB, UK), and approximately 50 ng of genomic DNA.

    Article Title: Genetic variants in the enhancer region of the thymidylate synthase gene in the Chilean population
    Article Snippet: Two oligonucleotide primers were synthesized specifically to amplify the polymorphic region of TSER (sense primer 5′-GTGGCTCCTGCGT TTCCCCC-3′ and antisense primer 5′-GCTCCGAGC CGG CCACAGGCATGGCGCGG-3′) [ ]. .. Amplification reactions were performed in a 25-µl volume containing 0.2 µg genomic DNA, 0.2 m m dNTPs , 0.5 µ m of each primer, 1.0 units of Taq polymerase (New England BioLabs, Ipswich, MA, USA) and 2.5 µl of 10 × Taq PCR buffer. .. Cycling conditions were: one cycle at 94 °C for 5 min with hot start; 30 cycles of 40 s at 94 °C, 1 min at 62 °C and 40 s at 72 °C; and one elongation cycle for 5 min at 72 °C.

    Article Title: Mechanisms of pseudosubstrate inhibition of the anaphase promoting complex by Acm1
    Article Snippet: The antisense primer annealed to URA3 and extended to the last amino acid of CDH1 . .. EP-PCR conditions were as follows: 2 μg/ml CDH1- Δ 200-URA3 -pAS2, 0.2 μM of each primer, 1 mM dTTP, 1 mM dCTP, 0.2 mM dATP, 0.2 mM dGTP, 6 mM MgCl2 , 50 μM MnCl2 , 1 × PCR buffer (−MgCl2 ) (Invitrogen) and 0.5 U/μl Taq polymerase (New England Biolabs). .. After 16 cycles of amplification, 1 ml of product was purified using a Qiagen PCR purification column (Qiagen, Inc.) and co-transformed into YJB1212 with ∼2.5 μg of gapped CDH1- Δ 200-URA3 -pAS2 vector.

    Article Title: Targeted gene inactivation in zebrafish using engineered zinc finger nucleases
    Article Snippet: Colonies from the desired selection plates were washed off the plates and their plasmid DNA was recovered as a pool as previously described . .. Each pool of individual fingers was amplified from the plasmid DNA by PCR: 50 μl reaction with 1 units NEB DNA Taq polymerase using 50 ng plasmid DNA template and 1 mM each primer ( ); denature for 1 min at 95°C, 28 cycles consisting of 94°C for 20 sec, 55°C for 20 sec and 72°C for 20 sec, followed by a final extension at 72°C for 6 min. .. The PCR products were run on a 1.5% agarose TAE gel, and the appropriate bands were excised and purified using a Qiagen PCR purification kit.

    Article Title: Pseudomonas aeruginosa AlgR Phosphorylation Modulates Rhamnolipid Production and Motility
    Article Snippet: Electrophoretic mobility shift assays (EMSAs) were performed using purified AlgR proteins, as described previously ( ). .. DNA fragments were prepared by PCR amplification of PAO1 chromosomal DNA with Taq polymerase (New England BioLabs) and the oligonucleotides listed in for fimU and rhlA . .. Oligonucleotides were 5′-end labeled with [γ-32 P]ATP (6,000 Ci/mmol) (Perkin-Elmer) with polynucleotide kinase (New England BioLabs).

    Article Title: Evidence for Acquisition of Legionella Type IV Secretion Substrates via Interdomain Horizontal Gene Transfer
    Article Snippet: Finally, we screened the individual plate by the same PCR technique to identify the mutant of interest. .. PCRs were performed using Taq polymerase (New England Biolabs, Massachusetts) with the following protocol: denaturation at 95°C for 5 min, followed by 35 cycles of denaturation at 95°C for 10 s, annealing at 68°C for 30 s, and amplification at 72°C for 3 min.

    Article Title: Identification of Quantitative Trait Loci and Candidate Genes for Maize Starch Granule Size through Association Mapping
    Article Snippet: DNA was extracted from seedlings of 26 maize lines with the largest starch granule sizes and 21 maize lines with the smallest starch granule sizes . .. PCR reaction mixes (20 µl) contained 1 µl of NEB (New England BioLabs Inc., Ipswich, MA, USA) Taq DNA Polymerase, 4 µl of 5× NEB PCR Buffer, 0.5 µl of dNTP mixture, 0.5 µl each of the two primers, and 1 µl of template DNA. .. The PCR reaction was carried out in a Bio-Rad Thermal cycler (Bio-Rad Laboratories, Inc., Hercules, CA, USA)with an initial denaturation at 94 °C for 3 min followed by 34 cycles of denaturation for 10 s at 94 °C, annealing for 1 min at 64 °C and extension for 1 min at 68 °C, with a final extension for 10 min at 68 °C.

    Crystallization Assay:

    Article Title: Post-crystallization improvement of RNA crystal diffraction quality
    Article Snippet: Taq DNA polymerase, 5,000 U/mL (New England Biolabs). .. Taq DNA polymerase, 5,000 U/mL (New England Biolabs).

    Selection:

    Article Title: Automated selection of aptamers against cholangiocarcinoma cells on an integrated microfluidic platform
    Article Snippet: After each round of Cell-SELEX, the magnetic field was activated such that the negative control cell-magnetic bead complexes could be trapped in the negative selection micropump/micromixer chamber. .. The PCR chamber was preloaded with 0.5 μ l of the forward primer (5′-GGCAGGAAGACAAACA-3′, 0.5 μ M), 0.5 μ l of the reverse primer (5′-ACAGCACCACAGACCA-3′, 0.5 μ M), 0.5 μ l of deoxynucleotide triphosphates (0.2 mM), 2 μ l of ssDNA (0.1 μ M) or the previous amplified product from the last round of Cell-SELEX, 0.125 μ l of Taq DNA polymerase (New England Biolabs, USA), and double-distilled water (ddH2 O) to a final volume of 25 μ l. The PCR was initiated at 95 °C for 10 min followed by 30 cycles of 95 °C for 30 s, 63 °C for 15 s, and 72 °C for 30 s. A final extension step at 72 °C for 10 min was also incorporated into the protocol.

    Article Title: Promoter Activation by CII, a Potent Transcriptional Activator from Bacteriophage 186
    Article Snippet: The resulting PCR product was used as template for error-prone PCR with Taq DNA polymerase (New England Biolabs) and MgCl2 supplemented to a total concentration of 6 m m . .. The product of error-prone PCR was subcloned back into pZS45-CII145 via BamHI and XhoI restriction sites and transformed into screening strain IM18 by electroporation.

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Paragraph title: Error-prone PCR and selection of feedback-inhibition-resistant tyrA mutants. ... Using the plasmid pZE21:: tyrA WT as template, 10, 20, and 30 amplification cycles with the primers mentioned above were performed using Taq DNA polymerase (New England Biolabs).

    Article Title: Targeted gene inactivation in zebrafish using engineered zinc finger nucleases
    Article Snippet: Colonies from the desired selection plates were washed off the plates and their plasmid DNA was recovered as a pool as previously described . .. Each pool of individual fingers was amplified from the plasmid DNA by PCR: 50 μl reaction with 1 units NEB DNA Taq polymerase using 50 ng plasmid DNA template and 1 mM each primer ( ); denature for 1 min at 95°C, 28 cycles consisting of 94°C for 20 sec, 55°C for 20 sec and 72°C for 20 sec, followed by a final extension at 72°C for 6 min.

    Electrophoretic Mobility Shift Assay:

    Article Title: Pseudomonas aeruginosa AlgR Phosphorylation Modulates Rhamnolipid Production and Motility
    Article Snippet: Electrophoretic mobility shift assays (EMSAs) were performed using purified AlgR proteins, as described previously ( ). .. DNA fragments were prepared by PCR amplification of PAO1 chromosomal DNA with Taq polymerase (New England BioLabs) and the oligonucleotides listed in for fimU and rhlA .

    Recombinant:

    Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
    Article Snippet: E. coli transformation, plasmid isolation, and recombinant techniques were performed using standard methods ( ). .. Restriction enzymes, T4 DNA ligase, and Taq DNA polymerase were purchased from NEB.

    Article Title: The Alternative Sigma Factor ?H Is Required for Toxin Gene Expression by Bacillus anthracis
    Article Snippet: Preparation of plasmid DNA from E. coli , transformation of E. coli , and recombinant DNA techniques were performed using standard procedures ( ). .. Restriction enzymes and T4 DNA ligase were purchased from Promega (Madison, WI) and Fisher Scientific, and Taq DNA polymerase was purchased from New England Biolabs (Beverly, MA).

    Nested PCR:

    Article Title: Field evaluation of a real time loop-mediated isothermal amplification assay (RealAmp) for malaria diagnosis in Cruzeiro do Sul, Acre, Brazil
    Article Snippet: Paragraph title: Nested PCR ... Reactions were performed in 20 μL total volume containing 1X buffer, 2.5 mM MgCl2 , 200 mM dNTPs, 200 nM primers, and 1.25 units of Taq Polymerase (New England Biolabs, Ipswich, MA).

    Two Hybrid Screening:

    Article Title: Mechanisms of pseudosubstrate inhibition of the anaphase promoting complex by Acm1
    Article Snippet: Paragraph title: EP-PCR mutagenesis of CDH1- Δ 200 and yeast two-hybrid screening ... EP-PCR conditions were as follows: 2 μg/ml CDH1- Δ 200-URA3 -pAS2, 0.2 μM of each primer, 1 mM dTTP, 1 mM dCTP, 0.2 mM dATP, 0.2 mM dGTP, 6 mM MgCl2 , 50 μM MnCl2 , 1 × PCR buffer (−MgCl2 ) (Invitrogen) and 0.5 U/μl Taq polymerase (New England Biolabs).

    Activated Clotting Time Assay:

    Article Title: Molecular Analysis of the Enterobacter sakazakii O-Antigen Gene Locus
    Article Snippet: The O:1 serotype-specific PCR used the forward primer ESO:1F (5′-CAC GTT CGC CCT GCA AAA AT-3′) and a reverse primer, ESO:1R (5′-GCA AGC GGC CAG ACT GGA TA-3′), designed from sequence information within wehC to generate a 341-bp amplicon. .. PCR amplification was performed with 50-μl volumes containing 1× PCR buffer containing 1.5 mM MgCl2 (New England Biolabs, Herts, United Kingdom), 100 mM deoxynucleoside triphosphates, 0.1 μM of each primer (MWG Biotech AG, Ebersberg, Germany), 2.5 U Taq DNA polymerase (New England Biolabs, Herts, United Kingdom), 100 ng template DNA, and PCR-grade water (Invitrogen, CA).

    Plasmid Preparation:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Nucleotide analogue mutagenesis was carried out in the presence of 2 and 20 μM 8-oxo-2′-deoxyguanosine (8-oxo-dGTP) and 6-(2-deoxy-β- d -ribofuranosyl)-3,4-dihydro-8H-pyrimido-(4,5-c)(1,2)oxazin-7-one (dPTP) ( ). .. Using the plasmid pZE21:: tyrA WT as template, 10, 20, and 30 amplification cycles with the primers mentioned above were performed using Taq DNA polymerase (New England Biolabs). .. The 1.1-kbp PCR products were gel purified, and the mutated tyrA genes were amplified in a second PCR under regular conditions.

    Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
    Article Snippet: Non-methylated plasmid DNA for electroporation into B. anthracis ( ; ) was obtained from E. coli GM2163 cells. .. Restriction enzymes, T4 DNA ligase, and Taq DNA polymerase were purchased from NEB.

    Article Title: The Alternative Sigma Factor ?H Is Required for Toxin Gene Expression by Bacillus anthracis
    Article Snippet: B. anthracis was electroporated with unmethylated plasmid DNA from E. coli GM2163 as described elsewhere ( ). .. Restriction enzymes and T4 DNA ligase were purchased from Promega (Madison, WI) and Fisher Scientific, and Taq DNA polymerase was purchased from New England Biolabs (Beverly, MA).

    Article Title: Mechanisms of pseudosubstrate inhibition of the anaphase promoting complex by Acm1
    Article Snippet: EP-PCR conditions were as follows: 2 μg/ml CDH1- Δ 200-URA3 -pAS2, 0.2 μM of each primer, 1 mM dTTP, 1 mM dCTP, 0.2 mM dATP, 0.2 mM dGTP, 6 mM MgCl2 , 50 μM MnCl2 , 1 × PCR buffer (−MgCl2 ) (Invitrogen) and 0.5 U/μl Taq polymerase (New England Biolabs). .. After 16 cycles of amplification, 1 ml of product was purified using a Qiagen PCR purification column (Qiagen, Inc.) and co-transformed into YJB1212 with ∼2.5 μg of gapped CDH1- Δ 200-URA3 -pAS2 vector.

    Article Title: Targeted gene inactivation in zebrafish using engineered zinc finger nucleases
    Article Snippet: Colonies from the desired selection plates were washed off the plates and their plasmid DNA was recovered as a pool as previously described . .. Each pool of individual fingers was amplified from the plasmid DNA by PCR: 50 μl reaction with 1 units NEB DNA Taq polymerase using 50 ng plasmid DNA template and 1 mM each primer ( ); denature for 1 min at 95°C, 28 cycles consisting of 94°C for 20 sec, 55°C for 20 sec and 72°C for 20 sec, followed by a final extension at 72°C for 6 min. .. The PCR products were run on a 1.5% agarose TAE gel, and the appropriate bands were excised and purified using a Qiagen PCR purification kit.

    Article Title: Pseudomonas aeruginosa AlgR Phosphorylation Modulates Rhamnolipid Production and Motility
    Article Snippet: DNA fragments were prepared by PCR amplification of PAO1 chromosomal DNA with Taq polymerase (New England BioLabs) and the oligonucleotides listed in for fimU and rhlA . .. DNA fragments were prepared by PCR amplification of PAO1 chromosomal DNA with Taq polymerase (New England BioLabs) and the oligonucleotides listed in for fimU and rhlA .

    Software:

    Article Title: Eco-Evolutionary Processes Generating Diversity Among Bottlenose Dolphin, Tursiops truncatus, Populations off Baja California, Mexico
    Article Snippet: The PCRs were performed in 25µL volumes consisting of 10 mM Tris–HCl, 50 mM KCl, 2.5 mM MgCl2 , 0.25 mM each dNTP, 0.12 µM each primer: L15812 (TRO): 5′ CCT CCC TAA GAC TCA AGG AAG 3′ and (D): 5′ CCT GAA GTA AGA ACC AGA TG 3′ (Rosel et al. ), 1.25 U of Taq DNA polymerase (NEB, UK), and approximately 50 ng of genomic DNA. .. PCR products were purified using purification spin columns (QIAGEN, UK) and then sequenced in an automatic sequencer (ABI 3730 Gene Analyzer, Applied Biosystems).

    Negative Control:

    Article Title: Automated selection of aptamers against cholangiocarcinoma cells on an integrated microfluidic platform
    Article Snippet: After each round of Cell-SELEX, the magnetic field was activated such that the negative control cell-magnetic bead complexes could be trapped in the negative selection micropump/micromixer chamber. .. The PCR chamber was preloaded with 0.5 μ l of the forward primer (5′-GGCAGGAAGACAAACA-3′, 0.5 μ M), 0.5 μ l of the reverse primer (5′-ACAGCACCACAGACCA-3′, 0.5 μ M), 0.5 μ l of deoxynucleotide triphosphates (0.2 mM), 2 μ l of ssDNA (0.1 μ M) or the previous amplified product from the last round of Cell-SELEX, 0.125 μ l of Taq DNA polymerase (New England Biolabs, USA), and double-distilled water (ddH2 O) to a final volume of 25 μ l. The PCR was initiated at 95 °C for 10 min followed by 30 cycles of 95 °C for 30 s, 63 °C for 15 s, and 72 °C for 30 s. A final extension step at 72 °C for 10 min was also incorporated into the protocol.

    Denaturing Gradient Gel Electrophoresis:

    Article Title: Growth of Dehalobacter and Dehalococcoides spp. during Degradation of Chlorinated Ethanes
    Article Snippet: PCR was performed in triplicate 50-μl reactions containing 1× ThermoPol PCR buffer (New England Biolabs, Mississauga, Ontario, Canada), 0.5 μM concentrations of each primer, 0.5 mM deoxynucleoside triphosphates, 1.5 U of Taq DNA polymerase (New Englands Biolabs), and 50 ng of DNA. .. Positive clones (42 clones) were sequenced by the University Health Network Research DNA Sequencing Facility (Toronto, Ontario, Canada) with the primer 27f, and then the sequence closest match was identified with the blastn utility of GenBank.

    Agarose Gel Electrophoresis:

    Article Title: Mechanisms of pseudosubstrate inhibition of the anaphase promoting complex by Acm1
    Article Snippet: EP-PCR conditions were as follows: 2 μg/ml CDH1- Δ 200-URA3 -pAS2, 0.2 μM of each primer, 1 mM dTTP, 1 mM dCTP, 0.2 mM dATP, 0.2 mM dGTP, 6 mM MgCl2 , 50 μM MnCl2 , 1 × PCR buffer (−MgCl2 ) (Invitrogen) and 0.5 U/μl Taq polymerase (New England Biolabs). .. After 16 cycles of amplification, 1 ml of product was purified using a Qiagen PCR purification column (Qiagen, Inc.) and co-transformed into YJB1212 with ∼2.5 μg of gapped CDH1- Δ 200-URA3 -pAS2 vector.

    In Vitro:

    Article Title: Direct Selection of RNA Beacon Aptamers
    Article Snippet: The original RNA pool was produced by in vitro transcription. .. Reactions contained 0.125 μM DM001, 1 μM DM003, 1 μM DM004 , 200 μM dNTPs, 1X buffer (provided with enzyme) and 12.5 U/mL Taq DNA polymerase (New England Biolabs).

    Spectrophotometry:

    Article Title: Molecular Analysis of the Enterobacter sakazakii O-Antigen Gene Locus
    Article Snippet: Total DNA was prepared using the Wizard Genomic DNA purification kit (Promega, Madison, WI) and quantified by spectrophotometry (NanoDrop, Wilmington, DE). .. PCR amplification was performed with 50-μl volumes containing 1× PCR buffer containing 1.5 mM MgCl2 (New England Biolabs, Herts, United Kingdom), 100 mM deoxynucleoside triphosphates, 0.1 μM of each primer (MWG Biotech AG, Ebersberg, Germany), 2.5 U Taq DNA polymerase (New England Biolabs, Herts, United Kingdom), 100 ng template DNA, and PCR-grade water (Invitrogen, CA).

    Produced:

    Article Title: Direct Selection of RNA Beacon Aptamers
    Article Snippet: The original RNA pool was produced by in vitro transcription. .. Reactions contained 0.125 μM DM001, 1 μM DM003, 1 μM DM004 , 200 μM dNTPs, 1X buffer (provided with enzyme) and 12.5 U/mL Taq DNA polymerase (New England Biolabs).

    Activation Assay:

    Article Title: Promoter Activation by CII, a Potent Transcriptional Activator from Bacteriophage 186
    Article Snippet: Paragraph title: Genetic Screen for CII Activation Mutants ... The resulting PCR product was used as template for error-prone PCR with Taq DNA polymerase (New England Biolabs) and MgCl2 supplemented to a total concentration of 6 m m .

    DNA Purification:

    Article Title: Molecular Analysis of the Enterobacter sakazakii O-Antigen Gene Locus
    Article Snippet: Total DNA was prepared using the Wizard Genomic DNA purification kit (Promega, Madison, WI) and quantified by spectrophotometry (NanoDrop, Wilmington, DE). .. PCR amplification was performed with 50-μl volumes containing 1× PCR buffer containing 1.5 mM MgCl2 (New England Biolabs, Herts, United Kingdom), 100 mM deoxynucleoside triphosphates, 0.1 μM of each primer (MWG Biotech AG, Ebersberg, Germany), 2.5 U Taq DNA polymerase (New England Biolabs, Herts, United Kingdom), 100 ng template DNA, and PCR-grade water (Invitrogen, CA).

    Gel Extraction:

    Article Title: Mechanisms of pseudosubstrate inhibition of the anaphase promoting complex by Acm1
    Article Snippet: EP-PCR conditions were as follows: 2 μg/ml CDH1- Δ 200-URA3 -pAS2, 0.2 μM of each primer, 1 mM dTTP, 1 mM dCTP, 0.2 mM dATP, 0.2 mM dGTP, 6 mM MgCl2 , 50 μM MnCl2 , 1 × PCR buffer (−MgCl2 ) (Invitrogen) and 0.5 U/μl Taq polymerase (New England Biolabs). .. After 16 cycles of amplification, 1 ml of product was purified using a Qiagen PCR purification column (Qiagen, Inc.) and co-transformed into YJB1212 with ∼2.5 μg of gapped CDH1- Δ 200-URA3 -pAS2 vector.

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    New England Biolabs taq dna polymerase supertaq
    <t>DNA</t> modification and bypass during bisulfite treatment. ( A ) Scheme of the chemical reaction of cytidine: Treatment with bisulfite generates the non-aromatic, non-planar 5,6-dihydrouridine-6-sulfonate (dhU6S), which decomposes to uracil upon treatment with base (and heat). ( B ) Primer extension activity of different polymerases <t>(Taq,</t> 5D4, 3A10, E10, TgoT) on template T1 either unmodified (C), bisulfite-treated and desulfonated (Reagent 1, 80°C, 20 min) (converting dC to dU) (D) or bisulfite-treated (Reagent 1) but not desulphonated (converting dC to dhU6S) (S). Polymerases 5D4 and 3A10 are able to generate full-length (+20) products even from the non-desulphonated template (S). ( C ) Time-course comparison of primer extension activity of Taq and 5D4 on T1 either unmodified (C) or bisulfite-treated with (D) or without desulphonation (S). (P: primer).
    Taq Dna Polymerase Supertaq, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    DNA modification and bypass during bisulfite treatment. ( A ) Scheme of the chemical reaction of cytidine: Treatment with bisulfite generates the non-aromatic, non-planar 5,6-dihydrouridine-6-sulfonate (dhU6S), which decomposes to uracil upon treatment with base (and heat). ( B ) Primer extension activity of different polymerases (Taq, 5D4, 3A10, E10, TgoT) on template T1 either unmodified (C), bisulfite-treated and desulfonated (Reagent 1, 80°C, 20 min) (converting dC to dU) (D) or bisulfite-treated (Reagent 1) but not desulphonated (converting dC to dhU6S) (S). Polymerases 5D4 and 3A10 are able to generate full-length (+20) products even from the non-desulphonated template (S). ( C ) Time-course comparison of primer extension activity of Taq and 5D4 on T1 either unmodified (C) or bisulfite-treated with (D) or without desulphonation (S). (P: primer).

    Journal: Nucleic Acids Research

    Article Title: A polymerase engineered for bisulfite sequencing

    doi: 10.1093/nar/gkv798

    Figure Lengend Snippet: DNA modification and bypass during bisulfite treatment. ( A ) Scheme of the chemical reaction of cytidine: Treatment with bisulfite generates the non-aromatic, non-planar 5,6-dihydrouridine-6-sulfonate (dhU6S), which decomposes to uracil upon treatment with base (and heat). ( B ) Primer extension activity of different polymerases (Taq, 5D4, 3A10, E10, TgoT) on template T1 either unmodified (C), bisulfite-treated and desulfonated (Reagent 1, 80°C, 20 min) (converting dC to dU) (D) or bisulfite-treated (Reagent 1) but not desulphonated (converting dC to dhU6S) (S). Polymerases 5D4 and 3A10 are able to generate full-length (+20) products even from the non-desulphonated template (S). ( C ) Time-course comparison of primer extension activity of Taq and 5D4 on T1 either unmodified (C) or bisulfite-treated with (D) or without desulphonation (S). (P: primer).

    Article Snippet: Taq DNA polymerase (SuperTaq) was obtained from HT Biotechnology (Cambridge) and Q5 polymerase from New England Biolabs.

    Techniques: Modification, Activity Assay

    Amplification of 24 individual human genomic loci. Comparison of PCR performance of Taq (top) panel, with two different Taq/5D4 blends (5/1 (middle panel); 10/1 (bottom panel)) on the amplification of 24 different promotor regions in bisulfite-treated and fully desulphonated human genomic DNA. Both blends are able to amplify a significantly larger number of loci than Taq alone and together enable amplification of 18 out of 24 loci (75%).

    Journal: Nucleic Acids Research

    Article Title: A polymerase engineered for bisulfite sequencing

    doi: 10.1093/nar/gkv798

    Figure Lengend Snippet: Amplification of 24 individual human genomic loci. Comparison of PCR performance of Taq (top) panel, with two different Taq/5D4 blends (5/1 (middle panel); 10/1 (bottom panel)) on the amplification of 24 different promotor regions in bisulfite-treated and fully desulphonated human genomic DNA. Both blends are able to amplify a significantly larger number of loci than Taq alone and together enable amplification of 18 out of 24 loci (75%).

    Article Snippet: Taq DNA polymerase (SuperTaq) was obtained from HT Biotechnology (Cambridge) and Q5 polymerase from New England Biolabs.

    Techniques: Amplification, Polymerase Chain Reaction

    PCR amplification of bisulfite-treated plasmid templates. PCR amplification of bisulfite-treated high GC and low GC content templates ranging from 200–600 bp in size using fully desulphonated templates and three different 5D4/Taq blends (1/10, 1/5, 1/1) with progressively lower Taq content and Taq alone on low dC content plasmid regions (top panel) and high dC content plasmid regions (bottom panel). On templates with low dC content (and hence lower levels of dU and residual dhU6S adducts post bisulfite treatment and desulphonation) either Taq or Taq/5D4 polymerase blends with a high amount of Taq perform best. In contrast on the higher dC content templates only blends containing 5D4 yield amplicons with Taq/5D4 blends (10/1; 5/1) superior to 5D4/Taq 1/1 blend, while Taq alone does not yield any amplification products. Thus only 5D4/Taq blends are able to copy the high GC content templates indicating that the blended enzymes are more efficient at copying templates containing sporadic dUs (and dhU6S adducts) and dU homopolymer stretches. Low molecular weight bands result from primer-dimer formation. (M: E-Gel ® Low Range Quantitative DNA Ladder).

    Journal: Nucleic Acids Research

    Article Title: A polymerase engineered for bisulfite sequencing

    doi: 10.1093/nar/gkv798

    Figure Lengend Snippet: PCR amplification of bisulfite-treated plasmid templates. PCR amplification of bisulfite-treated high GC and low GC content templates ranging from 200–600 bp in size using fully desulphonated templates and three different 5D4/Taq blends (1/10, 1/5, 1/1) with progressively lower Taq content and Taq alone on low dC content plasmid regions (top panel) and high dC content plasmid regions (bottom panel). On templates with low dC content (and hence lower levels of dU and residual dhU6S adducts post bisulfite treatment and desulphonation) either Taq or Taq/5D4 polymerase blends with a high amount of Taq perform best. In contrast on the higher dC content templates only blends containing 5D4 yield amplicons with Taq/5D4 blends (10/1; 5/1) superior to 5D4/Taq 1/1 blend, while Taq alone does not yield any amplification products. Thus only 5D4/Taq blends are able to copy the high GC content templates indicating that the blended enzymes are more efficient at copying templates containing sporadic dUs (and dhU6S adducts) and dU homopolymer stretches. Low molecular weight bands result from primer-dimer formation. (M: E-Gel ® Low Range Quantitative DNA Ladder).

    Article Snippet: Taq DNA polymerase (SuperTaq) was obtained from HT Biotechnology (Cambridge) and Q5 polymerase from New England Biolabs.

    Techniques: Polymerase Chain Reaction, Amplification, Plasmid Preparation, Gas Chromatography, Molecular Weight

    Degree of methylation of individual CpG sites. Promoter regions of four genes ( A — prkcdbp , B — dab2ip , C — ptgs2 , D — ezh2) were amplified with either Taq ot 5D4/Taq blends, using bisulfide-treated genomic DNA from normal cells or LNCapP cells as a template and subjected to deep sequencing. Cyan—methylated CpGs, orange—unmethylated CpGs. Individual CpGs are numbered starting from the 5′ end of the amplicon.

    Journal: Nucleic Acids Research

    Article Title: A polymerase engineered for bisulfite sequencing

    doi: 10.1093/nar/gkv798

    Figure Lengend Snippet: Degree of methylation of individual CpG sites. Promoter regions of four genes ( A — prkcdbp , B — dab2ip , C — ptgs2 , D — ezh2) were amplified with either Taq ot 5D4/Taq blends, using bisulfide-treated genomic DNA from normal cells or LNCapP cells as a template and subjected to deep sequencing. Cyan—methylated CpGs, orange—unmethylated CpGs. Individual CpGs are numbered starting from the 5′ end of the amplicon.

    Article Snippet: Taq DNA polymerase (SuperTaq) was obtained from HT Biotechnology (Cambridge) and Q5 polymerase from New England Biolabs.

    Techniques: Methylation, Amplification, Sequencing