tth dna polymerase Search Results


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  • 99
    Thermo Fisher r tth dna polymerase
    R Tth Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Millipore tth polymerase
    Tth Polymerase, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Boehringer Mannheim tth dna polymerase
    The analysis of <t>DNA</t> product synthesized by <t>Tth</t> and Δ Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.
    Tth Dna Polymerase, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 90/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Promega tth dna polymerase
    Reverse Transcriptase mediated ribosomal RNA (rRNA) depletion is most effective with AMV RT and requires all steps to be effective. Nucleic acids were eluted from FTA cards with midgut contents of An. gambiae that had been exposed to a bloodmeal containing West Nile virus (WNV) placed on them. RNA and <t>DNA</t> was then extracted to obtain total nucleic acid. The nucleic acid was then treated with DNase I and then purified to obtain total RNA. This RNA was then subjected to cDNA synthesis with a panel of reverse transcriptases (n = 3 for each treatment) in the presence (+probes) or absence (- probes) of DNA probes specific to rRNA. The RTs tested were <t>Tth</t> DNA polymerase, Superscript III (SSIII), Superscript IV (SSIV), AMV and MMLV. All the samples were then treated with RNase H and then DNase I to remove the RNA present in an RNA:DNA hybrid and cDNA, respectively. The samples were then purified and subjected to qRT-PCR with primer probe combinations specific for 18S rRNA (A), 28S rRNA (B) or WNV (C). Further tests were performed exclusively with AMV RT. Panels D and E show the results of qRT-PCR for samples that underwent the process of depletion but omitting some step or reagent. 18S (D) and 28S (E) rRNA was quantified in the input RNA, RNA with no RT added, RNA with no depletion probes added and RNA treated with RT with depletion probes. All statistical tests were performed by One-Way ANOVA with Tukey test for multiple comparisons. ****Indicates p-value
    Tth Dna Polymerase, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Toyobo tth dna polymerase
    Reverse Transcriptase mediated ribosomal RNA (rRNA) depletion is most effective with AMV RT and requires all steps to be effective. Nucleic acids were eluted from FTA cards with midgut contents of An. gambiae that had been exposed to a bloodmeal containing West Nile virus (WNV) placed on them. RNA and <t>DNA</t> was then extracted to obtain total nucleic acid. The nucleic acid was then treated with DNase I and then purified to obtain total RNA. This RNA was then subjected to cDNA synthesis with a panel of reverse transcriptases (n = 3 for each treatment) in the presence (+probes) or absence (- probes) of DNA probes specific to rRNA. The RTs tested were <t>Tth</t> DNA polymerase, Superscript III (SSIII), Superscript IV (SSIV), AMV and MMLV. All the samples were then treated with RNase H and then DNase I to remove the RNA present in an RNA:DNA hybrid and cDNA, respectively. The samples were then purified and subjected to qRT-PCR with primer probe combinations specific for 18S rRNA (A), 28S rRNA (B) or WNV (C). Further tests were performed exclusively with AMV RT. Panels D and E show the results of qRT-PCR for samples that underwent the process of depletion but omitting some step or reagent. 18S (D) and 28S (E) rRNA was quantified in the input RNA, RNA with no RT added, RNA with no depletion probes added and RNA treated with RT with depletion probes. All statistical tests were performed by One-Way ANOVA with Tukey test for multiple comparisons. ****Indicates p-value
    Tth Dna Polymerase, supplied by Toyobo, used in various techniques. Bioz Stars score: 90/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher recombinant tth dna polymerase xl
    Reverse Transcriptase mediated ribosomal RNA (rRNA) depletion is most effective with AMV RT and requires all steps to be effective. Nucleic acids were eluted from FTA cards with midgut contents of An. gambiae that had been exposed to a bloodmeal containing West Nile virus (WNV) placed on them. RNA and <t>DNA</t> was then extracted to obtain total nucleic acid. The nucleic acid was then treated with DNase I and then purified to obtain total RNA. This RNA was then subjected to cDNA synthesis with a panel of reverse transcriptases (n = 3 for each treatment) in the presence (+probes) or absence (- probes) of DNA probes specific to rRNA. The RTs tested were <t>Tth</t> DNA polymerase, Superscript III (SSIII), Superscript IV (SSIV), AMV and MMLV. All the samples were then treated with RNase H and then DNase I to remove the RNA present in an RNA:DNA hybrid and cDNA, respectively. The samples were then purified and subjected to qRT-PCR with primer probe combinations specific for 18S rRNA (A), 28S rRNA (B) or WNV (C). Further tests were performed exclusively with AMV RT. Panels D and E show the results of qRT-PCR for samples that underwent the process of depletion but omitting some step or reagent. 18S (D) and 28S (E) rRNA was quantified in the input RNA, RNA with no RT added, RNA with no depletion probes added and RNA treated with RT with depletion probes. All statistical tests were performed by One-Way ANOVA with Tukey test for multiple comparisons. ****Indicates p-value
    Recombinant Tth Dna Polymerase Xl, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Epicentre Biotechnologies masteramp tth dna polymerase
    Reverse Transcriptase mediated ribosomal RNA (rRNA) depletion is most effective with AMV RT and requires all steps to be effective. Nucleic acids were eluted from FTA cards with midgut contents of An. gambiae that had been exposed to a bloodmeal containing West Nile virus (WNV) placed on them. RNA and <t>DNA</t> was then extracted to obtain total nucleic acid. The nucleic acid was then treated with DNase I and then purified to obtain total RNA. This RNA was then subjected to cDNA synthesis with a panel of reverse transcriptases (n = 3 for each treatment) in the presence (+probes) or absence (- probes) of DNA probes specific to rRNA. The RTs tested were <t>Tth</t> DNA polymerase, Superscript III (SSIII), Superscript IV (SSIV), AMV and MMLV. All the samples were then treated with RNase H and then DNase I to remove the RNA present in an RNA:DNA hybrid and cDNA, respectively. The samples were then purified and subjected to qRT-PCR with primer probe combinations specific for 18S rRNA (A), 28S rRNA (B) or WNV (C). Further tests were performed exclusively with AMV RT. Panels D and E show the results of qRT-PCR for samples that underwent the process of depletion but omitting some step or reagent. 18S (D) and 28S (E) rRNA was quantified in the input RNA, RNA with no RT added, RNA with no depletion probes added and RNA treated with RT with depletion probes. All statistical tests were performed by One-Way ANOVA with Tukey test for multiple comparisons. ****Indicates p-value
    Masteramp Tth Dna Polymerase, supplied by Epicentre Biotechnologies, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Toyobo 1x tth dna polymerase buffer
    Reverse Transcriptase mediated ribosomal RNA (rRNA) depletion is most effective with AMV RT and requires all steps to be effective. Nucleic acids were eluted from FTA cards with midgut contents of An. gambiae that had been exposed to a bloodmeal containing West Nile virus (WNV) placed on them. RNA and <t>DNA</t> was then extracted to obtain total nucleic acid. The nucleic acid was then treated with DNase I and then purified to obtain total RNA. This RNA was then subjected to cDNA synthesis with a panel of reverse transcriptases (n = 3 for each treatment) in the presence (+probes) or absence (- probes) of DNA probes specific to rRNA. The RTs tested were <t>Tth</t> DNA polymerase, Superscript III (SSIII), Superscript IV (SSIV), AMV and MMLV. All the samples were then treated with RNase H and then DNase I to remove the RNA present in an RNA:DNA hybrid and cDNA, respectively. The samples were then purified and subjected to qRT-PCR with primer probe combinations specific for 18S rRNA (A), 28S rRNA (B) or WNV (C). Further tests were performed exclusively with AMV RT. Panels D and E show the results of qRT-PCR for samples that underwent the process of depletion but omitting some step or reagent. 18S (D) and 28S (E) rRNA was quantified in the input RNA, RNA with no RT added, RNA with no depletion probes added and RNA treated with RT with depletion probes. All statistical tests were performed by One-Way ANOVA with Tukey test for multiple comparisons. ****Indicates p-value
    1x Tth Dna Polymerase Buffer, supplied by Toyobo, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher recombinant thermostable thermus thermophilus tth dna polymerase
    Reverse Transcriptase mediated ribosomal RNA (rRNA) depletion is most effective with AMV RT and requires all steps to be effective. Nucleic acids were eluted from FTA cards with midgut contents of An. gambiae that had been exposed to a bloodmeal containing West Nile virus (WNV) placed on them. RNA and <t>DNA</t> was then extracted to obtain total nucleic acid. The nucleic acid was then treated with DNase I and then purified to obtain total RNA. This RNA was then subjected to cDNA synthesis with a panel of reverse transcriptases (n = 3 for each treatment) in the presence (+probes) or absence (- probes) of DNA probes specific to rRNA. The RTs tested were <t>Tth</t> DNA polymerase, Superscript III (SSIII), Superscript IV (SSIV), AMV and MMLV. All the samples were then treated with RNase H and then DNase I to remove the RNA present in an RNA:DNA hybrid and cDNA, respectively. The samples were then purified and subjected to qRT-PCR with primer probe combinations specific for 18S rRNA (A), 28S rRNA (B) or WNV (C). Further tests were performed exclusively with AMV RT. Panels D and E show the results of qRT-PCR for samples that underwent the process of depletion but omitting some step or reagent. 18S (D) and 28S (E) rRNA was quantified in the input RNA, RNA with no RT added, RNA with no depletion probes added and RNA treated with RT with depletion probes. All statistical tests were performed by One-Way ANOVA with Tukey test for multiple comparisons. ****Indicates p-value
    Recombinant Thermostable Thermus Thermophilus Tth Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Toyobo recombinant δ tth dna polymerase
    The analysis of <t>DNA</t> product synthesized by Tth and <t>Δ</t> Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.
    Recombinant δ Tth Dna Polymerase, supplied by Toyobo, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Toyobo r tth dna polymerase rt pcr high plus
    The analysis of <t>DNA</t> product synthesized by Tth and <t>Δ</t> Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.
    R Tth Dna Polymerase Rt Pcr High Plus, supplied by Toyobo, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Toyobo delta tth dna polymerase
    The analysis of <t>DNA</t> product synthesized by Tth and <t>Δ</t> Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.
    Delta Tth Dna Polymerase, supplied by Toyobo, used in various techniques. Bioz Stars score: 80/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Thermo Fisher super tth dna polymerase sphaeroq
    The analysis of <t>DNA</t> product synthesized by Tth and <t>Δ</t> Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.
    Super Tth Dna Polymerase Sphaeroq, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 80/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher thermostable recombinant tth dna polymerase
    The analysis of <t>DNA</t> product synthesized by Tth and <t>Δ</t> Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.
    Thermostable Recombinant Tth Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher thermostable recombinant tth rtth dna polymerase
    The analysis of <t>DNA</t> product synthesized by Tth and <t>Δ</t> Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.
    Thermostable Recombinant Tth Rtth Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher thermostable enzyme tth dna polymerase
    The analysis of <t>DNA</t> product synthesized by Tth and <t>Δ</t> Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.
    Thermostable Enzyme Tth Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Promega thermus thermophilus tth dna polymerase
    The analysis of <t>DNA</t> product synthesized by Tth and <t>Δ</t> Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.
    Thermus Thermophilus Tth Dna Polymerase, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The analysis of DNA product synthesized by Tth and Δ Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.

    Journal: Nucleic Acids Research

    Article Title: Elongation of repetitive DNA by DNA polymerase from a hyperthermophilic bacterium Thermus thermophilus

    doi:

    Figure Lengend Snippet: The analysis of DNA product synthesized by Tth and Δ Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.

    Article Snippet: The amount of DNA synthesized was 12 pmol by Tth DNA polymerase and < 1 pmol by Δ Tth DNA polymerase, a mutant Tth DNA polymerase lacking 5′→3′ exonuclease activity , when no oligoDNA was added to the reaction mixture (Table ).

    Techniques: Synthesized, Agarose Gel Electrophoresis, Sequencing

    Reverse Transcriptase mediated ribosomal RNA (rRNA) depletion is most effective with AMV RT and requires all steps to be effective. Nucleic acids were eluted from FTA cards with midgut contents of An. gambiae that had been exposed to a bloodmeal containing West Nile virus (WNV) placed on them. RNA and DNA was then extracted to obtain total nucleic acid. The nucleic acid was then treated with DNase I and then purified to obtain total RNA. This RNA was then subjected to cDNA synthesis with a panel of reverse transcriptases (n = 3 for each treatment) in the presence (+probes) or absence (- probes) of DNA probes specific to rRNA. The RTs tested were Tth DNA polymerase, Superscript III (SSIII), Superscript IV (SSIV), AMV and MMLV. All the samples were then treated with RNase H and then DNase I to remove the RNA present in an RNA:DNA hybrid and cDNA, respectively. The samples were then purified and subjected to qRT-PCR with primer probe combinations specific for 18S rRNA (A), 28S rRNA (B) or WNV (C). Further tests were performed exclusively with AMV RT. Panels D and E show the results of qRT-PCR for samples that underwent the process of depletion but omitting some step or reagent. 18S (D) and 28S (E) rRNA was quantified in the input RNA, RNA with no RT added, RNA with no depletion probes added and RNA treated with RT with depletion probes. All statistical tests were performed by One-Way ANOVA with Tukey test for multiple comparisons. ****Indicates p-value

    Journal: Virology

    Article Title: A reverse-transcription/RNase H based protocol for depletion of mosquito ribosomal RNA facilitates viral intrahost evolution analysis, transcriptomics and pathogen discovery

    doi: 10.1016/j.virol.2018.12.020

    Figure Lengend Snippet: Reverse Transcriptase mediated ribosomal RNA (rRNA) depletion is most effective with AMV RT and requires all steps to be effective. Nucleic acids were eluted from FTA cards with midgut contents of An. gambiae that had been exposed to a bloodmeal containing West Nile virus (WNV) placed on them. RNA and DNA was then extracted to obtain total nucleic acid. The nucleic acid was then treated with DNase I and then purified to obtain total RNA. This RNA was then subjected to cDNA synthesis with a panel of reverse transcriptases (n = 3 for each treatment) in the presence (+probes) or absence (- probes) of DNA probes specific to rRNA. The RTs tested were Tth DNA polymerase, Superscript III (SSIII), Superscript IV (SSIV), AMV and MMLV. All the samples were then treated with RNase H and then DNase I to remove the RNA present in an RNA:DNA hybrid and cDNA, respectively. The samples were then purified and subjected to qRT-PCR with primer probe combinations specific for 18S rRNA (A), 28S rRNA (B) or WNV (C). Further tests were performed exclusively with AMV RT. Panels D and E show the results of qRT-PCR for samples that underwent the process of depletion but omitting some step or reagent. 18S (D) and 28S (E) rRNA was quantified in the input RNA, RNA with no RT added, RNA with no depletion probes added and RNA treated with RT with depletion probes. All statistical tests were performed by One-Way ANOVA with Tukey test for multiple comparisons. ****Indicates p-value

    Article Snippet: For initial experiments, we tested a panel of reverse transcriptases (RTs), including Tth DNA polymerase (in the presence of Mn2 +, Promega), Superscript III (SSIII, ThermoFisher), Superscript IV (SSIV, ThermoFisher), Avian myeloblastosis virus (AMV, NEB) and Moloney Murine Leukemia Virus (MMLV, NEB).

    Techniques: Purification, Quantitative RT-PCR

    Incorporation of 3’-Aep-dCMP by commercially available A-family DNA polymerases (AF-DNAPs). ( A ) Top, the schematic representation of single 3’-Aep-dCMP incorporation by Taq, Tth, Tfl, BF, KF, or Bsu. Bottom, DNA fragment analysis of the primer (N) and the primer plus an incorporated 3’-Aep-dCMP by BF (N + 1). ( B ) Activities of single 3’-Aep-dCMP incorporation by Taq, Tth, Tfl, BF, KF, or Bsu, respectively. The primer-extension assays were performed as described in the Methods using 0.1, 0.2, 0.4, 0.8, 2, 4, 10, 20, or 40 μM of 3’-Aep-dCTP in the reaction.

    Journal: Scientific Reports

    Article Title: Enzymatic Cleavage of 3’-Esterified Nucleotides Enables a Long, Continuous DNA Synthesis

    doi: 10.1038/s41598-020-64541-z

    Figure Lengend Snippet: Incorporation of 3’-Aep-dCMP by commercially available A-family DNA polymerases (AF-DNAPs). ( A ) Top, the schematic representation of single 3’-Aep-dCMP incorporation by Taq, Tth, Tfl, BF, KF, or Bsu. Bottom, DNA fragment analysis of the primer (N) and the primer plus an incorporated 3’-Aep-dCMP by BF (N + 1). ( B ) Activities of single 3’-Aep-dCMP incorporation by Taq, Tth, Tfl, BF, KF, or Bsu, respectively. The primer-extension assays were performed as described in the Methods using 0.1, 0.2, 0.4, 0.8, 2, 4, 10, 20, or 40 μM of 3’-Aep-dCTP in the reaction.

    Article Snippet: Tfl and Tth DNA polymerases were purchased from Promega (Madison, WI).

    Techniques:

    The analysis of DNA product synthesized by Tth and Δ Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.

    Journal: Nucleic Acids Research

    Article Title: Elongation of repetitive DNA by DNA polymerase from a hyperthermophilic bacterium Thermus thermophilus

    doi:

    Figure Lengend Snippet: The analysis of DNA product synthesized by Tth and Δ Tth DNA polymerases by restriction enzymes. The DNA product synthesized in the standard reaction mixture at 74°C for 30 min by Tth or Δ Tth DNA polymerase in the presence of 1 µg/ml of the indicated oligoDNA (third line) was electrophoresed on an agarose gel under non-denaturing conditions before (lanes 1, 3, 5 and 7) or after (lanes 2, 4, 6 and 8) treatment with 1 U/ml of the restriction enzyme indicated at the bottom. The size markers are shown in kb on the left. The recognition sequence of the restriction enzyme is shown below each enzyme.

    Article Snippet: DNA synthesis by Tth DNA polymerase (Boehringer Mannheim, Mannheim, Germany) or recombinant Δ Tth DNA polymerase (Toyobo, Osaka, Japan), a mutant of Tth DNA polymerase in which 5′→3′ exonuclease activity is lacking , was carried out essentially as described previously ( ).

    Techniques: Synthesized, Agarose Gel Electrophoresis, Sequencing