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  • 99
    Thermo Fisher taq dna polymerase
    Taq Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 33236 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taq dna polymerase/product/Thermo Fisher
    Average 99 stars, based on 33236 article reviews
    Price from $9.99 to $1999.99
    taq dna polymerase - by Bioz Stars, 2020-04
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    99
    Thermo Fisher platinum taq dna polymerase
    Effect of the molecular beacon primer on PCR specificity and sensitivity. ( A ) The target was M.tuberculosis <t>DNA.</t> All reaction mixtures contained 0.5 µg of human placental DNA. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder), the prominent band is at 500 bp; lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, ∼1000 genome copies; lanes 3, 6, 9 and 12, 100 copies; lanes 4, 7, 10 and 13, 10 copies; lanes 2–7, platinum <t>Taq</t> DNA polymerase; lanes 8–13, normal Taq DNA polymerase. ( B ) Detection of M.tuberculosis DNA in clinical sample by the described technique. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder); lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, DNA sample from sputum; lanes 3, 6, 9 and 12, ∼1000 genome copies; lanes 4, 7, 10 and 13, ∼100 genome copies; lanes 2–7, platinum Taq DNA polymerase; lanes 8–13, normal Taq DNA polymerase.
    Platinum Taq Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 19903 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/platinum taq dna polymerase/product/Thermo Fisher
    Average 99 stars, based on 19903 article reviews
    Price from $9.99 to $1999.99
    platinum taq dna polymerase - by Bioz Stars, 2020-04
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    99
    Thermo Fisher platinum taq polymerase supermix
    Effect of the molecular beacon primer on PCR specificity and sensitivity. ( A ) The target was M.tuberculosis <t>DNA.</t> All reaction mixtures contained 0.5 µg of human placental DNA. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder), the prominent band is at 500 bp; lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, ∼1000 genome copies; lanes 3, 6, 9 and 12, 100 copies; lanes 4, 7, 10 and 13, 10 copies; lanes 2–7, platinum <t>Taq</t> DNA polymerase; lanes 8–13, normal Taq DNA polymerase. ( B ) Detection of M.tuberculosis DNA in clinical sample by the described technique. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder); lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, DNA sample from sputum; lanes 3, 6, 9 and 12, ∼1000 genome copies; lanes 4, 7, 10 and 13, ∼100 genome copies; lanes 2–7, platinum Taq DNA polymerase; lanes 8–13, normal Taq DNA polymerase.
    Platinum Taq Polymerase Supermix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/platinum taq polymerase supermix/product/Thermo Fisher
    Average 99 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    platinum taq polymerase supermix - by Bioz Stars, 2020-04
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    93
    Thermo Fisher taq platinum polymerase
    Effect of the molecular beacon primer on PCR specificity and sensitivity. ( A ) The target was M.tuberculosis <t>DNA.</t> All reaction mixtures contained 0.5 µg of human placental DNA. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder), the prominent band is at 500 bp; lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, ∼1000 genome copies; lanes 3, 6, 9 and 12, 100 copies; lanes 4, 7, 10 and 13, 10 copies; lanes 2–7, platinum <t>Taq</t> DNA polymerase; lanes 8–13, normal Taq DNA polymerase. ( B ) Detection of M.tuberculosis DNA in clinical sample by the described technique. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder); lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, DNA sample from sputum; lanes 3, 6, 9 and 12, ∼1000 genome copies; lanes 4, 7, 10 and 13, ∼100 genome copies; lanes 2–7, platinum Taq DNA polymerase; lanes 8–13, normal Taq DNA polymerase.
    Taq Platinum Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 279 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taq platinum polymerase/product/Thermo Fisher
    Average 93 stars, based on 279 article reviews
    Price from $9.99 to $1999.99
    taq platinum polymerase - by Bioz Stars, 2020-04
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    Thermo Fisher high fidelity platinum taq polymerase
    Effect of the molecular beacon primer on PCR specificity and sensitivity. ( A ) The target was M.tuberculosis <t>DNA.</t> All reaction mixtures contained 0.5 µg of human placental DNA. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder), the prominent band is at 500 bp; lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, ∼1000 genome copies; lanes 3, 6, 9 and 12, 100 copies; lanes 4, 7, 10 and 13, 10 copies; lanes 2–7, platinum <t>Taq</t> DNA polymerase; lanes 8–13, normal Taq DNA polymerase. ( B ) Detection of M.tuberculosis DNA in clinical sample by the described technique. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder); lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, DNA sample from sputum; lanes 3, 6, 9 and 12, ∼1000 genome copies; lanes 4, 7, 10 and 13, ∼100 genome copies; lanes 2–7, platinum Taq DNA polymerase; lanes 8–13, normal Taq DNA polymerase.
    High Fidelity Platinum Taq Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 599 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/high fidelity platinum taq polymerase/product/Thermo Fisher
    Average 99 stars, based on 599 article reviews
    Price from $9.99 to $1999.99
    high fidelity platinum taq polymerase - by Bioz Stars, 2020-04
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    99
    Thermo Fisher platinum taq buffer
    Effect of the molecular beacon primer on PCR specificity and sensitivity. ( A ) The target was M.tuberculosis <t>DNA.</t> All reaction mixtures contained 0.5 µg of human placental DNA. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder), the prominent band is at 500 bp; lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, ∼1000 genome copies; lanes 3, 6, 9 and 12, 100 copies; lanes 4, 7, 10 and 13, 10 copies; lanes 2–7, platinum <t>Taq</t> DNA polymerase; lanes 8–13, normal Taq DNA polymerase. ( B ) Detection of M.tuberculosis DNA in clinical sample by the described technique. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder); lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, DNA sample from sputum; lanes 3, 6, 9 and 12, ∼1000 genome copies; lanes 4, 7, 10 and 13, ∼100 genome copies; lanes 2–7, platinum Taq DNA polymerase; lanes 8–13, normal Taq DNA polymerase.
    Platinum Taq Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 281 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/platinum taq buffer/product/Thermo Fisher
    Average 99 stars, based on 281 article reviews
    Price from $9.99 to $1999.99
    platinum taq buffer - by Bioz Stars, 2020-04
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    Effect of the molecular beacon primer on PCR specificity and sensitivity. ( A ) The target was M.tuberculosis DNA. All reaction mixtures contained 0.5 µg of human placental DNA. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder), the prominent band is at 500 bp; lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, ∼1000 genome copies; lanes 3, 6, 9 and 12, 100 copies; lanes 4, 7, 10 and 13, 10 copies; lanes 2–7, platinum Taq DNA polymerase; lanes 8–13, normal Taq DNA polymerase. ( B ) Detection of M.tuberculosis DNA in clinical sample by the described technique. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder); lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, DNA sample from sputum; lanes 3, 6, 9 and 12, ∼1000 genome copies; lanes 4, 7, 10 and 13, ∼100 genome copies; lanes 2–7, platinum Taq DNA polymerase; lanes 8–13, normal Taq DNA polymerase.

    Journal: Nucleic Acids Research

    Article Title: PCR hot start using primers with the structure of molecular beacons (hairpin-like structure)

    doi:

    Figure Lengend Snippet: Effect of the molecular beacon primer on PCR specificity and sensitivity. ( A ) The target was M.tuberculosis DNA. All reaction mixtures contained 0.5 µg of human placental DNA. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder), the prominent band is at 500 bp; lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, ∼1000 genome copies; lanes 3, 6, 9 and 12, 100 copies; lanes 4, 7, 10 and 13, 10 copies; lanes 2–7, platinum Taq DNA polymerase; lanes 8–13, normal Taq DNA polymerase. ( B ) Detection of M.tuberculosis DNA in clinical sample by the described technique. Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder); lanes 2–4 and 8–10, reverse primer as molecular beacon; lanes 5–7 and 11–13, linear reverse primer; lanes 2, 5, 8 and 11, DNA sample from sputum; lanes 3, 6, 9 and 12, ∼1000 genome copies; lanes 4, 7, 10 and 13, ∼100 genome copies; lanes 2–7, platinum Taq DNA polymerase; lanes 8–13, normal Taq DNA polymerase.

    Article Snippet: Platinum Taq DNA polymerase (complexed with anti- Taq antibodies; Life Technologies) was used as a control.

    Techniques: Polymerase Chain Reaction, DNA Gel Electrophoresis, Amplification, Marker, Nucleic Acid Electrophoresis

    Effect of the molecular beacon primer on the PCR specificity and sensitivity. The target was human DNA (exon 4 of p53 gene). Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder), the prominent band is 500 bp; lanes 2, 3 and 6, 7, reverse primer as molecular beacon; lanes 4, 5 and 8, 9, linear reverse primer; lanes 2, 4, 6 and 8, ∼1000 genome copies; lanes 3, 5, 7 and 9, ∼100 genome copies; lanes 2–5, platinum Taq DNA polymerase; lanes 8 and 9, usual Taq DNA polymerase. The platinum Taq DNA polymerase gave good amplification from 100 000+ genome copies (not shown).

    Journal: Nucleic Acids Research

    Article Title: PCR hot start using primers with the structure of molecular beacons (hairpin-like structure)

    doi:

    Figure Lengend Snippet: Effect of the molecular beacon primer on the PCR specificity and sensitivity. The target was human DNA (exon 4 of p53 gene). Gel electrophoresis in 2% agarose of the amplification products. Lane 1, DNA size marker (100 bp ladder), the prominent band is 500 bp; lanes 2, 3 and 6, 7, reverse primer as molecular beacon; lanes 4, 5 and 8, 9, linear reverse primer; lanes 2, 4, 6 and 8, ∼1000 genome copies; lanes 3, 5, 7 and 9, ∼100 genome copies; lanes 2–5, platinum Taq DNA polymerase; lanes 8 and 9, usual Taq DNA polymerase. The platinum Taq DNA polymerase gave good amplification from 100 000+ genome copies (not shown).

    Article Snippet: Platinum Taq DNA polymerase (complexed with anti- Taq antibodies; Life Technologies) was used as a control.

    Techniques: Polymerase Chain Reaction, Nucleic Acid Electrophoresis, Amplification, Marker

    Detection of the beta-lactamase gene in commercial Taq polymerase. Four dilutions of Amplitaq DNA polymerase were tested with primers for the beta-lactamase gene in the presence of 10 3 pUC19 plasmids (labeled 1000 pUC19 genomes) or H 2 O.

    Journal: PLoS ONE

    Article Title: Optimizing Taq Polymerase Concentration for Improved Signal-to-Noise in the Broad Range Detection of Low Abundance Bacteria

    doi: 10.1371/journal.pone.0007010

    Figure Lengend Snippet: Detection of the beta-lactamase gene in commercial Taq polymerase. Four dilutions of Amplitaq DNA polymerase were tested with primers for the beta-lactamase gene in the presence of 10 3 pUC19 plasmids (labeled 1000 pUC19 genomes) or H 2 O.

    Article Snippet: Taq polymerases The following DNA polymerases from commercial vendors designed for qPCR were used in the experiments reported here: Amplitaq Gold (ABI, CA; Roche lot # J02913); Platinum Taq (Invitrogen, CA; cat # 10966–026; lot 1169610); Platinum HiFi Taq (Invitrogen, CA; cat# 11304–011; lot# 1267490); HotStar Taq (Qiagen, CA; Mat # 1007837; lot # 124125007); JumpStart Taq (Sigma, MO; cat # D-6558; lot # 71K9029).

    Techniques: Labeling

    cul3 Testis but Not cul3 Soma Can Restore Caspase Activation and Spermatid Individualization to cul3 Testis Null Mutants (A) Schematic structure of the rescue constructs for cul3 Testis −/− male sterile flies. The constructs tr-cul3 Testis and tr-cul3 Soma are composed of the cul3 Testis isoform's promoter region (dark blue, consists of the intronic sequences flanked by exons 2 and 1D) and 5′ UTR (light blue) that were fused upstream of the coding regions (ORFs) of either cul3 Testis or cul3 Soma followed by the 3′ UTR of cul3 Testis . (B and C) Transcriptional expression from the transgenes was confirmed by RT–PCR analyses on RNA from testes of the indicated genotypes. The relative locations of the primers are indicated with black arrows in (A). “RT+Taq” and “Taq” indicate reactions with reverse transcriptase or without it, respectively, to control for possible genomic DNA contamination. (B) To differentiate between the cul3 Testis endogenous (endog.) and transgenic (transg.) cDNAs, we cleaved the RT-PCR fragments with XhoI, a unique restriction site in the transgene. Note that the RT-PCR product from cul3 mds1 ; tr-cul3 Testis but not from WT testes was cleaved by XhoI, confirming its transgenic source. (C) Transgenic expression of cul3 Soma ( tr - cul3 Soma ) in adult testis. Note the absence of the endogenous cul3 Testis cDNA band and in contrast, the presence of the transgenic cul3 Soma band in cul3 mds1 ; tr - cul3 Soma /+ testes. (D–I) Testes stained for cleaved caspase-3 (CM1, green) and spermatid's tail and ICs (phalloidin, red). (D) Mutant spermatids for cul3 Testis ( cul3 mds1 −/− ) manifest a block in caspase activation and spermatid individualization. (E) Either one or (F) two copies of transgenic cul3 Testis ( tr-cul3 Testis ) restores caspase activation, spermatid individualization, and fertility of cul3 mds1 −/− male flies. (G) Wild-type control testes. Note the CBs and WBs (green oval structures). (H–I) In contrast, dramatically reduced CM1-positive cysts are found in cul3 mds1 mutants, which ectopically express (H) one or (I) two copies of the cul3 Soma transgene ( tr-cul3 Soma ). These spermatids failed to individualize, no CBs and WBs are detected and the males are sterile. Scale bars 200 μm.

    Journal: PLoS Biology

    Article Title: A Ubiquitin Ligase Complex Regulates Caspase Activation During Sperm Differentiation in DrosophilaA Protein Complex Restrains a Homicidal Enzyme during Sperm Differentiation

    doi: 10.1371/journal.pbio.0050251

    Figure Lengend Snippet: cul3 Testis but Not cul3 Soma Can Restore Caspase Activation and Spermatid Individualization to cul3 Testis Null Mutants (A) Schematic structure of the rescue constructs for cul3 Testis −/− male sterile flies. The constructs tr-cul3 Testis and tr-cul3 Soma are composed of the cul3 Testis isoform's promoter region (dark blue, consists of the intronic sequences flanked by exons 2 and 1D) and 5′ UTR (light blue) that were fused upstream of the coding regions (ORFs) of either cul3 Testis or cul3 Soma followed by the 3′ UTR of cul3 Testis . (B and C) Transcriptional expression from the transgenes was confirmed by RT–PCR analyses on RNA from testes of the indicated genotypes. The relative locations of the primers are indicated with black arrows in (A). “RT+Taq” and “Taq” indicate reactions with reverse transcriptase or without it, respectively, to control for possible genomic DNA contamination. (B) To differentiate between the cul3 Testis endogenous (endog.) and transgenic (transg.) cDNAs, we cleaved the RT-PCR fragments with XhoI, a unique restriction site in the transgene. Note that the RT-PCR product from cul3 mds1 ; tr-cul3 Testis but not from WT testes was cleaved by XhoI, confirming its transgenic source. (C) Transgenic expression of cul3 Soma ( tr - cul3 Soma ) in adult testis. Note the absence of the endogenous cul3 Testis cDNA band and in contrast, the presence of the transgenic cul3 Soma band in cul3 mds1 ; tr - cul3 Soma /+ testes. (D–I) Testes stained for cleaved caspase-3 (CM1, green) and spermatid's tail and ICs (phalloidin, red). (D) Mutant spermatids for cul3 Testis ( cul3 mds1 −/− ) manifest a block in caspase activation and spermatid individualization. (E) Either one or (F) two copies of transgenic cul3 Testis ( tr-cul3 Testis ) restores caspase activation, spermatid individualization, and fertility of cul3 mds1 −/− male flies. (G) Wild-type control testes. Note the CBs and WBs (green oval structures). (H–I) In contrast, dramatically reduced CM1-positive cysts are found in cul3 mds1 mutants, which ectopically express (H) one or (I) two copies of the cul3 Soma transgene ( tr-cul3 Soma ). These spermatids failed to individualize, no CBs and WBs are detected and the males are sterile. Scale bars 200 μm.

    Article Snippet: The RNA was stored in −80 °C or immediately used for RT-PCR reactions using the SuperScriptTM III One-Step RT–PCR System with Platinums Taq DNA polymerase (Invitrogen).

    Techniques: Activation Assay, Construct, Expressing, Reverse Transcription Polymerase Chain Reaction, Transgenic Assay, Staining, Mutagenesis, Blocking Assay

    Reportable range for T. cruzi and triatomine intestine unit quantification by real-time qPCR. Multiplex Taq Man qPCR assays were carried out with serially diluted DNA extracted from reconstituted triatomine intestine samples containing T. cruzi epimastigotes, ranging from 10 5 to 0.5 T. cruzi equivalents ( a ) and 5 to 0.002 triatomine intestine unit equivalents ( b ). The slope, R 2 and amplification efficiency (Eff) are indicated in the chart

    Journal: Parasites & Vectors

    Article Title: Development of conventional and real-time multiplex PCR-based assays for estimation of natural infection rates and Trypanosoma cruzi load in triatomine vectors

    doi: 10.1186/s13071-017-2343-x

    Figure Lengend Snippet: Reportable range for T. cruzi and triatomine intestine unit quantification by real-time qPCR. Multiplex Taq Man qPCR assays were carried out with serially diluted DNA extracted from reconstituted triatomine intestine samples containing T. cruzi epimastigotes, ranging from 10 5 to 0.5 T. cruzi equivalents ( a ) and 5 to 0.002 triatomine intestine unit equivalents ( b ). The slope, R 2 and amplification efficiency (Eff) are indicated in the chart

    Article Snippet: Multiplex conventional PCR (cPCR) Conventional PCR assays were carried out in a final volume of 50 μl, containing: 5 μl DNA (20–25 ng), 5 μl 10× Taq Platinum buffer, 0.2 mM dNTPs, 4.5 mM MgCl2 , 1.25 U Taq Platinum DNA polymerase (Life Technologies, Carlsbad, CA, USA), 200 nM 121/122 primers (T. cruzi kDNA) [ , ] and 100 nM P2B/P6R primers (triatomine 12S rRNA gene).

    Techniques: Real-time Polymerase Chain Reaction, Multiplex Assay, Amplification

    MNN2 gene amplification efficiency on modified strain colonies picked from Petri dishes or micro-plate cultures. A ) 3 independent assays carried out on 23 colonies for each condition. Dark grey: Petri dishes cultures, light grey: liquid cultures. B ) Average of the 3 assays presented in A (standard deviation=19.55). C ) PCR product visualized on a 1% agarose gel stained with SYBR safe. Left panel (1): Petri dishes, right panel (2): micro-plate cultures, ML: Molecular ladder. All amplifications have been carried out with the Platinium Taq.

    Journal: BMC Research Notes

    Article Title: PCR on yeast colonies: an improved method for glyco-engineered Saccharomyces cerevisiae

    doi: 10.1186/1756-0500-6-201

    Figure Lengend Snippet: MNN2 gene amplification efficiency on modified strain colonies picked from Petri dishes or micro-plate cultures. A ) 3 independent assays carried out on 23 colonies for each condition. Dark grey: Petri dishes cultures, light grey: liquid cultures. B ) Average of the 3 assays presented in A (standard deviation=19.55). C ) PCR product visualized on a 1% agarose gel stained with SYBR safe. Left panel (1): Petri dishes, right panel (2): micro-plate cultures, ML: Molecular ladder. All amplifications have been carried out with the Platinium Taq.

    Article Snippet: With the Platinium Taq DNA polymerase (Invitrogen), the PCR mix was performed with final concentrations of: 1x manufacturer-supplied buffer, 1.5 mM MgCl2 , 0.2 mM dNTPs, 0.3 mM primers, 2.4% DMSO, 0.5 U of the Hot Start enzyme, H2 O to 20 μL.

    Techniques: Amplification, Modification, Standard Deviation, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining

    Efficiency of MNN5 gene amplification from wild type and modified strains. A ) Percentage of colonies picked from Petri dishes that amplified MNN5. The experiment was carried out on 47 colonies for each strain. Dark grey: BY4742; light grey: YiMMOgène. Amplification was carried out using the DreamTaq or the Platinium Taq. B ). PCR product visualisation, after amplification with the Platinium Taq, on a 1% agarose gel stained with SYBR safe. Left panel (1): BY4742, right panel (2): YiMMOgène, ML: Molecular ladder.

    Journal: BMC Research Notes

    Article Title: PCR on yeast colonies: an improved method for glyco-engineered Saccharomyces cerevisiae

    doi: 10.1186/1756-0500-6-201

    Figure Lengend Snippet: Efficiency of MNN5 gene amplification from wild type and modified strains. A ) Percentage of colonies picked from Petri dishes that amplified MNN5. The experiment was carried out on 47 colonies for each strain. Dark grey: BY4742; light grey: YiMMOgène. Amplification was carried out using the DreamTaq or the Platinium Taq. B ). PCR product visualisation, after amplification with the Platinium Taq, on a 1% agarose gel stained with SYBR safe. Left panel (1): BY4742, right panel (2): YiMMOgène, ML: Molecular ladder.

    Article Snippet: With the Platinium Taq DNA polymerase (Invitrogen), the PCR mix was performed with final concentrations of: 1x manufacturer-supplied buffer, 1.5 mM MgCl2 , 0.2 mM dNTPs, 0.3 mM primers, 2.4% DMSO, 0.5 U of the Hot Start enzyme, H2 O to 20 μL.

    Techniques: Amplification, Modification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining

    Efficiency of MNN5 gene amplification from liquid culture of modified strain. A ) Percentage of MNN5 amplification from 47 colonies picked on Petri dishes and 47 colonies picked from liquid cultures in micro-plates. B ) Growth curve in micro-plate (YPD medium), average of two colonies of YiMMOgène. The OD 600 was measured in a micro-plate reader. ♦: OD 600 after 6 hour culture (early exponential phase). ●: OD 600 after 17 hour culture (late exponential phase). ▄: OD 600 after 24 hour culture (early stationary phase). C ) Percentage amplification of MNN5 from 47 YiMMOgène after different culture times in micro-plate. All amplifications have been carried out with the Platinium Taq.

    Journal: BMC Research Notes

    Article Title: PCR on yeast colonies: an improved method for glyco-engineered Saccharomyces cerevisiae

    doi: 10.1186/1756-0500-6-201

    Figure Lengend Snippet: Efficiency of MNN5 gene amplification from liquid culture of modified strain. A ) Percentage of MNN5 amplification from 47 colonies picked on Petri dishes and 47 colonies picked from liquid cultures in micro-plates. B ) Growth curve in micro-plate (YPD medium), average of two colonies of YiMMOgène. The OD 600 was measured in a micro-plate reader. ♦: OD 600 after 6 hour culture (early exponential phase). ●: OD 600 after 17 hour culture (late exponential phase). ▄: OD 600 after 24 hour culture (early stationary phase). C ) Percentage amplification of MNN5 from 47 YiMMOgène after different culture times in micro-plate. All amplifications have been carried out with the Platinium Taq.

    Article Snippet: With the Platinium Taq DNA polymerase (Invitrogen), the PCR mix was performed with final concentrations of: 1x manufacturer-supplied buffer, 1.5 mM MgCl2 , 0.2 mM dNTPs, 0.3 mM primers, 2.4% DMSO, 0.5 U of the Hot Start enzyme, H2 O to 20 μL.

    Techniques: Amplification, Modification

    MNN5 gene amplification efficiency on modified strain colonies picked from Petri dishes or micro-plate cultures. A ) 3 independent assays carried out on 47 colonies for each condition. Dark grey: Petri dishes cultures, light grey: micro-plate cultures. B ) Average of the 3 assays presented in A (standard deviation =22.07). C) PCR product visualised on a 1% agarose gel stained with SYBR safe. Left panel (1): Petri dishes, right panel (2): micro-plate cultures, ML: Molecular ladder. All amplifications were carried out using Platinium Taq.

    Journal: BMC Research Notes

    Article Title: PCR on yeast colonies: an improved method for glyco-engineered Saccharomyces cerevisiae

    doi: 10.1186/1756-0500-6-201

    Figure Lengend Snippet: MNN5 gene amplification efficiency on modified strain colonies picked from Petri dishes or micro-plate cultures. A ) 3 independent assays carried out on 47 colonies for each condition. Dark grey: Petri dishes cultures, light grey: micro-plate cultures. B ) Average of the 3 assays presented in A (standard deviation =22.07). C) PCR product visualised on a 1% agarose gel stained with SYBR safe. Left panel (1): Petri dishes, right panel (2): micro-plate cultures, ML: Molecular ladder. All amplifications were carried out using Platinium Taq.

    Article Snippet: With the Platinium Taq DNA polymerase (Invitrogen), the PCR mix was performed with final concentrations of: 1x manufacturer-supplied buffer, 1.5 mM MgCl2 , 0.2 mM dNTPs, 0.3 mM primers, 2.4% DMSO, 0.5 U of the Hot Start enzyme, H2 O to 20 μL.

    Techniques: Amplification, Modification, Standard Deviation, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining

    PAGE analysis of primer extension experiments with single OXP-modified and PDE primers. Primer extension with Klenow fragment of DNA polymerase I of nonheated ( A ) and preheated ( B ) single OXP-modified reverse primer, respectively along template 2. The extension reactions were incubated at 25°C for the indicated times after which the reaction mixtures were quenched and analyzed. ( C ) Primer extension with Taq DNA polymerase of PDE and OXP forward primers (nonheated control and preheated sample) along template oligonucleotide 1. Extension reactions were incubated at 25°C for 15 min, after which the aliquots from reaction mixtures were quenched and analyzed.

    Journal: Nucleic Acids Research

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance

    doi: 10.1093/nar/gkn575

    Figure Lengend Snippet: PAGE analysis of primer extension experiments with single OXP-modified and PDE primers. Primer extension with Klenow fragment of DNA polymerase I of nonheated ( A ) and preheated ( B ) single OXP-modified reverse primer, respectively along template 2. The extension reactions were incubated at 25°C for the indicated times after which the reaction mixtures were quenched and analyzed. ( C ) Primer extension with Taq DNA polymerase of PDE and OXP forward primers (nonheated control and preheated sample) along template oligonucleotide 1. Extension reactions were incubated at 25°C for 15 min, after which the aliquots from reaction mixtures were quenched and analyzed.

    Article Snippet: Thermus aquaticus (Taq ) DNA polymerase (recombinant) and Platinum® Taq DNA Polymerases were purchased from Invitrogen (Carlsbad, CA, USA).

    Techniques: Polyacrylamide Gel Electrophoresis, Modification, Incubation

    Comparison of the performance of OXP-modified primers to other Hot Start DNA polymerases. ( A ) Agarose gel analysis of the PCR products resulting from the 35 thermal cycles of amplification of five copies of a 365-bp fragment from the HIV-1 tat gene using 0.5 μM unmodified, single OXP-modified and double OXP-modified primers. Reactions containing unmodified primers were amplified by Taq DNA polymerase, Platinum® Taq DNA Polymerase, AmpliTaq Gold® DNA Polymerase, HotStart-IT™ Taq DNA Polymerase and DyNazyme™ II Hot Start DNA Polymerase. Reactions containing single and double OXP-modified primers were amplified by Taq DNA polymerase. ( B ) Graphical representation of PCR amplicon yield. The results from triplicate experiments were averaged and are normalized to the yield of reactions containing single OXP-modified primers plus Taq DNA polymerase. Error bars represent the SD. (*), indicates Hot Start DNA polymerases.

    Journal: Nucleic Acids Research

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance

    doi: 10.1093/nar/gkn575

    Figure Lengend Snippet: Comparison of the performance of OXP-modified primers to other Hot Start DNA polymerases. ( A ) Agarose gel analysis of the PCR products resulting from the 35 thermal cycles of amplification of five copies of a 365-bp fragment from the HIV-1 tat gene using 0.5 μM unmodified, single OXP-modified and double OXP-modified primers. Reactions containing unmodified primers were amplified by Taq DNA polymerase, Platinum® Taq DNA Polymerase, AmpliTaq Gold® DNA Polymerase, HotStart-IT™ Taq DNA Polymerase and DyNazyme™ II Hot Start DNA Polymerase. Reactions containing single and double OXP-modified primers were amplified by Taq DNA polymerase. ( B ) Graphical representation of PCR amplicon yield. The results from triplicate experiments were averaged and are normalized to the yield of reactions containing single OXP-modified primers plus Taq DNA polymerase. Error bars represent the SD. (*), indicates Hot Start DNA polymerases.

    Article Snippet: Thermus aquaticus (Taq ) DNA polymerase (recombinant) and Platinum® Taq DNA Polymerases were purchased from Invitrogen (Carlsbad, CA, USA).

    Techniques: Modification, Agarose Gel Electrophoresis, Polymerase Chain Reaction, Amplification

    Dependence of pre-PCR heating of PTE on primer dimer accumulation. Agarose gel analysis of primer dimer accumulation with preheated single OXP-modified primers in nontemplate system. Mixture of both primers was preheated at 95°C in 1× PCR buffer (pH 8.4 at 25°C) for increasing amounts of time. Samples were cooled on ice water, the Taq DNA polymerase was added followed by PCR amplification. PCR cycle parameters: 95°C (2 min); 30 cycles of [95°C (40 s); 56°C (30 s); and 72°C (2 min)]; 72°C (7 min). Primer dimer amplicon, indicated on the gel image, ran as a 50–80 bp DNA fragment (left lane: 50-bp ladder).

    Journal: Nucleic Acids Research

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance

    doi: 10.1093/nar/gkn575

    Figure Lengend Snippet: Dependence of pre-PCR heating of PTE on primer dimer accumulation. Agarose gel analysis of primer dimer accumulation with preheated single OXP-modified primers in nontemplate system. Mixture of both primers was preheated at 95°C in 1× PCR buffer (pH 8.4 at 25°C) for increasing amounts of time. Samples were cooled on ice water, the Taq DNA polymerase was added followed by PCR amplification. PCR cycle parameters: 95°C (2 min); 30 cycles of [95°C (40 s); 56°C (30 s); and 72°C (2 min)]; 72°C (7 min). Primer dimer amplicon, indicated on the gel image, ran as a 50–80 bp DNA fragment (left lane: 50-bp ladder).

    Article Snippet: Thermus aquaticus (Taq ) DNA polymerase (recombinant) and Platinum® Taq DNA Polymerases were purchased from Invitrogen (Carlsbad, CA, USA).

    Techniques: Polymerase Chain Reaction, Agarose Gel Electrophoresis, Modification, Amplification

    Evaluation of thermolabile primers in one-step RT–PCR. For each gene of interest ( PBGD , ABCA-1 and β-actin ), the PCR primers were unmodified, single OXP-modified or double OXP-modified primers. RT utilized a polydT 18 primer. Reactions contained Taq DNA polymerase and M-MLV reverse transcriptase.

    Journal: Nucleic Acids Research

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance

    doi: 10.1093/nar/gkn575

    Figure Lengend Snippet: Evaluation of thermolabile primers in one-step RT–PCR. For each gene of interest ( PBGD , ABCA-1 and β-actin ), the PCR primers were unmodified, single OXP-modified or double OXP-modified primers. RT utilized a polydT 18 primer. Reactions contained Taq DNA polymerase and M-MLV reverse transcriptase.

    Article Snippet: Thermus aquaticus (Taq ) DNA polymerase (recombinant) and Platinum® Taq DNA Polymerases were purchased from Invitrogen (Carlsbad, CA, USA).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Modification

    Effect of PEITC on RASSF1A promoter demethylation. LNCaP cells (3.5 × 10 4 cells/ml) were seeded in 10-cm tissue culture plates. LNCaP cells were treated with 0.01% DMSO, PEITC (2.5 or 5 μM), or 5-Aza (2.5 μM) and TSA (0.5 μM) for 5 days. The media containing the drugs was changed on day 3 and day 5. Cells were harvested on day 6 for all the experiments. A) Representative images from the methylation-specific PCR experiments. Genomic DNA was extracted from cells harvested on day 6, and the bisulfite conversion was subsequently performed. M: methylated, U: unmethylated. The relative intensity of the methylated and unmethylated bands was measured using ImageJ software, and the values are presented in the lower panel. B) The methylation pattern of the 16 CpG sites in the fragment of RASSF1A promoter was analyzed using bisulfite genomic sequencing. Solid circles indicate methylated CpG sites, and empty circles indicate unmethylated CpG sites. Ten representative clones from each of the 3 independent experiments were selected for analysis. C) The percentage of CpG methylation is presented. The methylation percentage was calculated from three independent experiments as the number of methylated CpG sites over the total number of CpG sites examined. D) The effect of PEITC on RASSF1A mRNA expression. Total mRNA was isolated and analyzed using quantitative real-time PCR. The data are presented as the mean ± SEM of 3 independent experiments. * P

    Journal: Pharmacological research

    Article Title: Epigenetic Reactivation of RASSF1A by Phenethyl Isothiocyanate (PEITC) and promotion of apoptosis in LNCaP cells

    doi: 10.1016/j.phrs.2016.10.021

    Figure Lengend Snippet: Effect of PEITC on RASSF1A promoter demethylation. LNCaP cells (3.5 × 10 4 cells/ml) were seeded in 10-cm tissue culture plates. LNCaP cells were treated with 0.01% DMSO, PEITC (2.5 or 5 μM), or 5-Aza (2.5 μM) and TSA (0.5 μM) for 5 days. The media containing the drugs was changed on day 3 and day 5. Cells were harvested on day 6 for all the experiments. A) Representative images from the methylation-specific PCR experiments. Genomic DNA was extracted from cells harvested on day 6, and the bisulfite conversion was subsequently performed. M: methylated, U: unmethylated. The relative intensity of the methylated and unmethylated bands was measured using ImageJ software, and the values are presented in the lower panel. B) The methylation pattern of the 16 CpG sites in the fragment of RASSF1A promoter was analyzed using bisulfite genomic sequencing. Solid circles indicate methylated CpG sites, and empty circles indicate unmethylated CpG sites. Ten representative clones from each of the 3 independent experiments were selected for analysis. C) The percentage of CpG methylation is presented. The methylation percentage was calculated from three independent experiments as the number of methylated CpG sites over the total number of CpG sites examined. D) The effect of PEITC on RASSF1A mRNA expression. Total mRNA was isolated and analyzed using quantitative real-time PCR. The data are presented as the mean ± SEM of 3 independent experiments. * P

    Article Snippet: Briefly, the DNA sequences were amplified by mixing bisulfite-converted DNA (500 ng) with primer MU379 (50 pmoles) and primer ML730 (50 pmoles) in reaction buffer (20 μl) containing dNTPs (200 μM each dNTP) using Platinum PCR Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA).

    Techniques: Methylation, Polymerase Chain Reaction, Software, Genomic Sequencing, Clone Assay, CpG Methylation Assay, Expressing, Isolation, Real-time Polymerase Chain Reaction