t7 dna polymerase Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    New England Biolabs t7 dna polymerase
    E. coli SSB proteins stimulate <t>T7</t> DNA polymerase progression across 5 ′ -CAG60 repeats. A , schematic diagram of assay. The star indicates 32 P-labeled primer. B , sequencing gel showing the primer extension kinetics of T7 DNA polymerase across
    T7 Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 500 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 dna polymerase/product/New England Biolabs
    Average 99 stars, based on 500 article reviews
    Price from $9.99 to $1999.99
    t7 dna polymerase - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    94
    Thermo Fisher t7 dna polymerase buffer
    Fig. 4. ( A ) In a homopolymer RT assay (polyA RNA, oligo-dT primer), L1 ORF2p produces RT products far in excess of the molecular weight of the template, indicative of template switching activity; AMV RT does not. ( B ) TPRT of various RNA species. The end-point of the L1 and Alu RNAs are indicated with the dotted line (not to scale). RNA number 4 has 38 nt of vector RNA after the polyA tail. URA3, fragment of S . cerevisiae URA3 RNA with and without a polyA tail. ( C ) Distribution of cDNA initiation points. Shown below each histogram is a full-length cDNA (from 3′ to 5′), with the positions of the reverse-transcribed polyG and polyA tracts indicated by ‘C n ’ and ‘TTTTTT’, respectively. Position zero marks the end of L1 sequence and the beginning of the polyA tail. The actual positions of the cDNA initiation are plotted in the histogram grouped into 10 nt bins. For the first cDNA (generated from RNA number 1 in B), many cDNAs end beyond the designed position at nucleotide 14 due to the addition of extra A residues to the RNA by <t>T7</t> polymerase (see Materials and methods). Transposition of RNA number 4 (the second cDNA from the top) yields two distinct populations of cDNA initiation points. Mutation of either the polyG or the polyA sequence (histograms three and four, respectively) removes the bias towards internal initiation of cDNA synthesis, although only the polyA mutation results in a statistically significant difference. Student’s two-tailed t -test comparison of wild type versus mutant, bins –20–30 with bins 30–60; polyG p -value = 0.17; polyA p -value = 0.05. For all three cDNAs from 3′ extended transcripts, all initiation points in the 50–60 bin occurred at nucleotide 53, the end of the transcript. The following number of highly truncated (endpoint
    T7 Dna Polymerase Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 98 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 dna polymerase buffer/product/Thermo Fisher
    Average 94 stars, based on 98 article reviews
    Price from $9.99 to $1999.99
    t7 dna polymerase buffer - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    91
    GE Healthcare t7 dna polymerase
    Use of PNAs to facilitate formation of an active primer–template complex and subsequent strand elongation by modified <t>T7</t> DNA polymerase.
    T7 Dna Polymerase, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 91/100, based on 233 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 dna polymerase/product/GE Healthcare
    Average 91 stars, based on 233 article reviews
    Price from $9.99 to $1999.99
    t7 dna polymerase - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    91
    Stratagene t7 dna polymerase
    Use of PNAs to facilitate formation of an active primer–template complex and subsequent strand elongation by modified <t>T7</t> DNA polymerase.
    T7 Dna Polymerase, supplied by Stratagene, used in various techniques. Bioz Stars score: 91/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 dna polymerase/product/Stratagene
    Average 91 stars, based on 40 article reviews
    Price from $9.99 to $1999.99
    t7 dna polymerase - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    91
    Roche t7 dna polymerase
    Use of PNAs to facilitate formation of an active primer–template complex and subsequent strand elongation by modified <t>T7</t> DNA polymerase.
    T7 Dna Polymerase, supplied by Roche, used in various techniques. Bioz Stars score: 91/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 dna polymerase/product/Roche
    Average 91 stars, based on 25 article reviews
    Price from $9.99 to $1999.99
    t7 dna polymerase - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    91
    TaKaRa t7 dna polymerase
    Use of PNAs to facilitate formation of an active primer–template complex and subsequent strand elongation by modified <t>T7</t> DNA polymerase.
    T7 Dna Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 dna polymerase/product/TaKaRa
    Average 91 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    t7 dna polymerase - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    91
    Promega t7 dna polymerase
    Use of PNAs to facilitate formation of an active primer–template complex and subsequent strand elongation by modified <t>T7</t> DNA polymerase.
    T7 Dna Polymerase, supplied by Promega, used in various techniques. Bioz Stars score: 91/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 dna polymerase/product/Promega
    Average 91 stars, based on 75 article reviews
    Price from $9.99 to $1999.99
    t7 dna polymerase - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    93
    Thermo Fisher t7pol
    Use of PNAs to facilitate formation of an active primer–template complex and subsequent strand elongation by modified <t>T7</t> DNA polymerase.
    T7pol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7pol/product/Thermo Fisher
    Average 93 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    t7pol - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    85
    GE Healthcare t7 dna polymerase kit
    Use of PNAs to facilitate formation of an active primer–template complex and subsequent strand elongation by modified <t>T7</t> DNA polymerase.
    T7 Dna Polymerase Kit, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 85/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 dna polymerase kit/product/GE Healthcare
    Average 85 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    t7 dna polymerase kit - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    Image Search Results


    E. coli SSB proteins stimulate T7 DNA polymerase progression across 5 ′ -CAG60 repeats. A , schematic diagram of assay. The star indicates 32 P-labeled primer. B , sequencing gel showing the primer extension kinetics of T7 DNA polymerase across

    Journal:

    Article Title: Single-stranded DNA-binding Protein in Vitro Eliminates the Orientation-dependent Impediment to Polymerase Passage on CAG/CTG Repeats *

    doi: 10.1074/jbc.M800153200

    Figure Lengend Snippet: E. coli SSB proteins stimulate T7 DNA polymerase progression across 5 ′ -CAG60 repeats. A , schematic diagram of assay. The star indicates 32 P-labeled primer. B , sequencing gel showing the primer extension kinetics of T7 DNA polymerase across

    Article Snippet: T7 exonuclease; N. AlwI nicking enzyme; EcoRI, NdeI, and XbaI restriction endonucleases; T7 DNA polymerase; and T4 polynucleotide kinase were from New England Biolabs. ϕ29 DNA polymerase was either a gift from Dr. M. Salas or purchased from New England Biolabs.

    Techniques: Labeling, Sequencing

    Kunkel mutagenesis. Single-stranded D NA with uracil incorporated (dU-ssDNA; green ( gray in the print version) above) is isolated from phage produced in CJ236 cells. Degenerate primers ( red ( black in the print version)) are annealed (mutagenesis regions indicated by asterices ) to the dU-ssDNA. T7 DNA polymerase and T4 DNA ligase and dNTPs are added to synthesize the complementary strand based on the dU-ssDNA template. This covalently closed circular double-stranded DNA (CCC-dsDNA) is then electroporated into E. coli SS320 cells, in which the uracil-containing template is deactivated and the new, mutagenic strand is preferentially replicated.

    Journal: Methods in enzymology

    Article Title: Protein and Antibody Engineering by Phage Display

    doi: 10.1016/bs.mie.2016.05.005

    Figure Lengend Snippet: Kunkel mutagenesis. Single-stranded D NA with uracil incorporated (dU-ssDNA; green ( gray in the print version) above) is isolated from phage produced in CJ236 cells. Degenerate primers ( red ( black in the print version)) are annealed (mutagenesis regions indicated by asterices ) to the dU-ssDNA. T7 DNA polymerase and T4 DNA ligase and dNTPs are added to synthesize the complementary strand based on the dU-ssDNA template. This covalently closed circular double-stranded DNA (CCC-dsDNA) is then electroporated into E. coli SS320 cells, in which the uracil-containing template is deactivated and the new, mutagenic strand is preferentially replicated.

    Article Snippet: 3,3′,5,5′-Tetramethylbenzidine (TMB) (Sigma, St. Louis, MO, catalog #: T0440) 0.5 M H2 SO4 Anti-M13-HRP conjugated antibody (GE Healthcare, Marlborough, MA, catalog #: GE27-9421-01) T7 DNA Polymerase (NEB, Ipswich, MA, catalog #: M0274L) T4 DNA Ligase (Invitrogen, Carlsbad, CA, catalog #: 15224-017) 25 m M dNTPs (Fisher, Hampton, NH, catalog #: FERR1121) 25 mg/mL Uridine (Sigma, St. Louis, MO, catalog #: U3750-100G) BugBuster 10X Protein Extraction Reagent (Millipore, Billerica, MA, catalog #: 70921) QIAquick PCR Purification Kit (Qiagen, Hilden, Germany, catalog #: 28104) QIAquick Gel Extraction Kit (Qiagen, Hilden, Germany, catalog #: 28704) QIAprep Spin Miniprep Kit (Qiagen, Hilden, Germany, catalog #: 27104) NucleoBond Xtra Maxi Plus (Macherey-Nagel, Düren, Germany, catalog #: 740416.50) Ni-NTA Agarose (Qiagen, Hilden, Germany, catalog #: 30210) DNaseI (Invitrogen, Carlsbad, CA, catalog #: 18047-019) SIGMAFast EDTA-free protease cocktail inhibitor (Sigma, St. Louis, MO, catalog #: S8830) Precision Plus Protein Dual Color Standards (BioRad, Hercules, CA, catalog #: 161-03741) Anti-His (C-term)-HRP antibody (Invitrogen, Carlsbad, CA, catalog #: 46-0707) Protein A Agarose (Pierce, Thermo Fisher Scientific, Waltham, MA, catalog #: 15918-014) Phusion High Fidelity PCR Master Mix with HF Buffer (NEB, Ipswich, MA, catalog #: M0531S) Restriction enzymes: BssHII, BsiWI, NheI, NsiI, FseI (NEB, Ipswich, MA, catalog #s: R0199S, R0553S, R0131S, R0127S, R0558S, respectively) Alkaline Phosphatase, Calf Intestinal (NEB, Ipswich, MA, catalog #: M0290S) Quick Ligation Kit (NEB, Ipswich, MA, catalog #: M2200S) Polyethylenimine (PEI) Linear, 25,000 MW (Polysciences, Warrington, PA, catalog #: 23966) FreeStyle™ 293 Expression Medium (ThermoFisher; Invitrogen, Carlsbad, CA, catalog #: 12338-018) M2 antibody (Sigma-Aldrich, St. Louis, MO, catalog #: F1804) Protein A-HRP conjugate (Invitrogen, Carlsbad, CA, catalog #: 101023)

    Techniques: Mutagenesis, Isolation, Produced, Countercurrent Chromatography

    Helicase displacement of a single nucleosome. ( a ) Experimental configuration. A single dsDNA molecule was unwound by a T7 helicase as the two strands of the DNA were held under 12 pN of force by an optical trap, which assisted helicase unwinding but was insufficient to mechanically separate the dsDNA ( Supplementary Fig. 1B ). The nucleosomal DNA template is specified in Supplementary Fig. 2A , Supplementary Table 1 and Methods. ( b ) Representative helicase-unwinding traces on a nucleosomal (black) or naked (grey) template. Helicase unwinding was interrupted by discrete pauses along the DNA template. Dashed lines indicate the dyad locations of the initial positioned nucleosome and the transferred nucleosome. N =49 traces. ( c ) Histogram of nucleosome transfer distance. A transfer distance was obtained from the first transfer event of each trace as indicated by the arrow in Fig. 2b . The histogram was obtained by pooling data from 49 traces. The prediction (not a fit) from the DNA looping model is plotted for comparison. The resulting Pearson test gives a reduced χ 2 of 0.31 with a P value of 0.95 (Methods; Supplementary Fig. 4A ).

    Journal: Nature Communications

    Article Title: DNA looping mediates nucleosome transfer

    doi: 10.1038/ncomms13337

    Figure Lengend Snippet: Helicase displacement of a single nucleosome. ( a ) Experimental configuration. A single dsDNA molecule was unwound by a T7 helicase as the two strands of the DNA were held under 12 pN of force by an optical trap, which assisted helicase unwinding but was insufficient to mechanically separate the dsDNA ( Supplementary Fig. 1B ). The nucleosomal DNA template is specified in Supplementary Fig. 2A , Supplementary Table 1 and Methods. ( b ) Representative helicase-unwinding traces on a nucleosomal (black) or naked (grey) template. Helicase unwinding was interrupted by discrete pauses along the DNA template. Dashed lines indicate the dyad locations of the initial positioned nucleosome and the transferred nucleosome. N =49 traces. ( c ) Histogram of nucleosome transfer distance. A transfer distance was obtained from the first transfer event of each trace as indicated by the arrow in Fig. 2b . The histogram was obtained by pooling data from 49 traces. The prediction (not a fit) from the DNA looping model is plotted for comparison. The resulting Pearson test gives a reduced χ 2 of 0.31 with a P value of 0.95 (Methods; Supplementary Fig. 4A ).

    Article Snippet: Replisomes were formed by pre-incubating 1 unit per μl T7 DNA polymerase (NEB) and 1 μM T7 helicase in reaction buffer on ice for 5 min, and then were added to a final concentration of 0.1 unit per μl T7 DNA polymerase (NEB) and 100 nM T7 helicase and incubated at 37 °C for 10 min.

    Techniques:

    Idling-turnover kinetics. An excess concentration of linear P/T (1 μM) formed by annealing oligonucleotide E to F was incubated with wild-type HSV-1 pol (40 nM, •), wild-type HSV-1 pol/UL42 complex (40 nM, ▪), T7 DNA polymerase holoenzyme (15 nM, ▴), Klenow fragment (20 nM, □), or HSV-1 D368A pol (20 nM, ○) in reaction buffer containing 300 μM dATP and [α- 32 ). The accumulation of radiolabeled dAMP was monitored by thin-layer chromatography, normalized to enzyme concentration, and plotted as a function of time. Apparent rate constants were estimated from the linear portion of each plot (—) by least-squares regression analysis.

    Journal: Journal of Virology

    Article Title: 3? to 5? Exonuclease Activity of Herpes Simplex Virus Type 1 DNA Polymerase Modulates Its Strand Displacement Activity

    doi: 10.1128/JVI.77.18.10147-10153.2003

    Figure Lengend Snippet: Idling-turnover kinetics. An excess concentration of linear P/T (1 μM) formed by annealing oligonucleotide E to F was incubated with wild-type HSV-1 pol (40 nM, •), wild-type HSV-1 pol/UL42 complex (40 nM, ▪), T7 DNA polymerase holoenzyme (15 nM, ▴), Klenow fragment (20 nM, □), or HSV-1 D368A pol (20 nM, ○) in reaction buffer containing 300 μM dATP and [α- 32 ). The accumulation of radiolabeled dAMP was monitored by thin-layer chromatography, normalized to enzyme concentration, and plotted as a function of time. Apparent rate constants were estimated from the linear portion of each plot (—) by least-squares regression analysis.

    Article Snippet: As a means for comparison, we followed the strand displacement rates for wild-type T7 DNA polymerase holoenzyme (New England Biolabs, Beverly, Mass.) or an exonuclease-deficient mutant T7 DNA polymerase holoenzyme (Sequenase version 2.0; USB, Inc., Cleveland, Ohio).

    Techniques: Concentration Assay, Incubation, Thin Layer Chromatography

    Fig. 4. ( A ) In a homopolymer RT assay (polyA RNA, oligo-dT primer), L1 ORF2p produces RT products far in excess of the molecular weight of the template, indicative of template switching activity; AMV RT does not. ( B ) TPRT of various RNA species. The end-point of the L1 and Alu RNAs are indicated with the dotted line (not to scale). RNA number 4 has 38 nt of vector RNA after the polyA tail. URA3, fragment of S . cerevisiae URA3 RNA with and without a polyA tail. ( C ) Distribution of cDNA initiation points. Shown below each histogram is a full-length cDNA (from 3′ to 5′), with the positions of the reverse-transcribed polyG and polyA tracts indicated by ‘C n ’ and ‘TTTTTT’, respectively. Position zero marks the end of L1 sequence and the beginning of the polyA tail. The actual positions of the cDNA initiation are plotted in the histogram grouped into 10 nt bins. For the first cDNA (generated from RNA number 1 in B), many cDNAs end beyond the designed position at nucleotide 14 due to the addition of extra A residues to the RNA by T7 polymerase (see Materials and methods). Transposition of RNA number 4 (the second cDNA from the top) yields two distinct populations of cDNA initiation points. Mutation of either the polyG or the polyA sequence (histograms three and four, respectively) removes the bias towards internal initiation of cDNA synthesis, although only the polyA mutation results in a statistically significant difference. Student’s two-tailed t -test comparison of wild type versus mutant, bins –20–30 with bins 30–60; polyG p -value = 0.17; polyA p -value = 0.05. For all three cDNAs from 3′ extended transcripts, all initiation points in the 50–60 bin occurred at nucleotide 53, the end of the transcript. The following number of highly truncated (endpoint

    Journal: The EMBO Journal

    Article Title: Human L1 element target-primed reverse transcription in vitro

    doi: 10.1093/emboj/cdf592

    Figure Lengend Snippet: Fig. 4. ( A ) In a homopolymer RT assay (polyA RNA, oligo-dT primer), L1 ORF2p produces RT products far in excess of the molecular weight of the template, indicative of template switching activity; AMV RT does not. ( B ) TPRT of various RNA species. The end-point of the L1 and Alu RNAs are indicated with the dotted line (not to scale). RNA number 4 has 38 nt of vector RNA after the polyA tail. URA3, fragment of S . cerevisiae URA3 RNA with and without a polyA tail. ( C ) Distribution of cDNA initiation points. Shown below each histogram is a full-length cDNA (from 3′ to 5′), with the positions of the reverse-transcribed polyG and polyA tracts indicated by ‘C n ’ and ‘TTTTTT’, respectively. Position zero marks the end of L1 sequence and the beginning of the polyA tail. The actual positions of the cDNA initiation are plotted in the histogram grouped into 10 nt bins. For the first cDNA (generated from RNA number 1 in B), many cDNAs end beyond the designed position at nucleotide 14 due to the addition of extra A residues to the RNA by T7 polymerase (see Materials and methods). Transposition of RNA number 4 (the second cDNA from the top) yields two distinct populations of cDNA initiation points. Mutation of either the polyG or the polyA sequence (histograms three and four, respectively) removes the bias towards internal initiation of cDNA synthesis, although only the polyA mutation results in a statistically significant difference. Student’s two-tailed t -test comparison of wild type versus mutant, bins –20–30 with bins 30–60; polyG p -value = 0.17; polyA p -value = 0.05. For all three cDNAs from 3′ extended transcripts, all initiation points in the 50–60 bin occurred at nucleotide 53, the end of the transcript. The following number of highly truncated (endpoint

    Article Snippet: L1 3′ RNA was produced by in vitro T7 polymerase-driven transcription of nucleotides 5672–6036 of L1.3 from pQF325 linearized with Bsa I. T7 transcription was performed for 1.5 h in 50 µl of buffer containing 2 µg DNA template, 40 mM Tris pH 7.5, 20 mM MgCl2 , 50 µg/ml bovine serum albumin, 500 µM each NTP, 10 mM DTT and 100 U T7 polymerase (Life Technologies) and 40 U of RNAsin (Promega).

    Techniques: Molecular Weight, Activity Assay, Plasmid Preparation, Sequencing, Generated, Mutagenesis, Two Tailed Test

    Use of PNAs to facilitate formation of an active primer–template complex and subsequent strand elongation by modified T7 DNA polymerase.

    Journal: Nucleic Acids Research

    Article Title: Strand invasion by mixed base PNAs and a PNA-peptide chimera

    doi:

    Figure Lengend Snippet: Use of PNAs to facilitate formation of an active primer–template complex and subsequent strand elongation by modified T7 DNA polymerase.

    Article Snippet: The labeling mix consisting of modified T7 DNA polymerase (Sequenase; US Biochemical, Cleveland, OH) (1 U/reaction) and [35 S]dATP (Amersham) were added and DNA sequencing using bound peptide–oligonucleotide as primer was carried out.

    Techniques: Modification