hotstart taq pcr  (Qiagen)


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    Structured Review

    Qiagen hotstart taq pcr
    Hotstart Taq Pcr, supplied by Qiagen, used in various techniques. Bioz Stars score: 87/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hotstart taq pcr/product/Qiagen
    Average 87 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    hotstart taq pcr - by Bioz Stars, 2020-04
    87/100 stars

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    Nested PCR:

    Article Title: Surface protein mutations in chronic hepatitis B patients who received hepatitis B vaccine therapy
    Article Snippet: .. A nested PCR was carried out in 100 µl of a mixture containing 5 µl of DNA using HotStart Taq PCR (Qiagen, Hilden, Germany). .. 5 µl extracted HBV DNA and 1 µl of the first round amplicon were used as template for the first and the second round PCR reactions, respectively.

    Polymerase Chain Reaction:

    Article Title: Surface protein mutations in chronic hepatitis B patients who received hepatitis B vaccine therapy
    Article Snippet: .. A nested PCR was carried out in 100 µl of a mixture containing 5 µl of DNA using HotStart Taq PCR (Qiagen, Hilden, Germany). .. 5 µl extracted HBV DNA and 1 µl of the first round amplicon were used as template for the first and the second round PCR reactions, respectively.

    Agarose Gel Electrophoresis:

    Article Title: Surface protein mutations in chronic hepatitis B patients who received hepatitis B vaccine therapy
    Article Snippet: A nested PCR was carried out in 100 µl of a mixture containing 5 µl of DNA using HotStart Taq PCR (Qiagen, Hilden, Germany). .. Finally, 3 µl of second- round PCR product was analyzed by electrophoresis in 1% agarose gel, stained with ethidium bromide, and visualized under ultraviolet light.

    Electrophoresis:

    Article Title: Surface protein mutations in chronic hepatitis B patients who received hepatitis B vaccine therapy
    Article Snippet: A nested PCR was carried out in 100 µl of a mixture containing 5 µl of DNA using HotStart Taq PCR (Qiagen, Hilden, Germany). .. Finally, 3 µl of second- round PCR product was analyzed by electrophoresis in 1% agarose gel, stained with ethidium bromide, and visualized under ultraviolet light.

    Staining:

    Article Title: Surface protein mutations in chronic hepatitis B patients who received hepatitis B vaccine therapy
    Article Snippet: A nested PCR was carried out in 100 µl of a mixture containing 5 µl of DNA using HotStart Taq PCR (Qiagen, Hilden, Germany). .. Finally, 3 µl of second- round PCR product was analyzed by electrophoresis in 1% agarose gel, stained with ethidium bromide, and visualized under ultraviolet light.

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    Qiagen hotstart taq dna polymerase
    Effect of individual constituents of the PCR on background fluorescence in the presence or absence of CABS. Each column represents the result of three PCRs performed: 1, in the absence of <t>Taq</t> Gold polymerase, but in the presence of oligonucleotides and template; 2, in the absence of Taq Gold polymerase and template, but in the presence of oligonucleotides; 3, in the absence of Taq Gold polymerase, template, and one oligonucleotide but in the presence of one oligonucleotide; 4, in the absence of primers and template, but in the presence of Taq Gold polymerase; 5, in the absence of template, but in the presence of both primers and polymerase; 6, as in case 3, except in the presence of the alternative primer of the primer combination; 7, in which specific amplicon is produced; and 8, in which no amplicon is expected. Conditions 1, 2, and 6 show the effect of CABS in reducing the background fluorescence caused by the single-strand oligonucleotides. The reaction with CABS in conditions 5 and 8 shows the residual effect caused by primer/dimers, in the case of 8 with additional template <t>DNA.</t>
    Hotstart Taq Dna Polymerase, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hotstart taq dna polymerase/product/Qiagen
    Average 99 stars, based on 110 article reviews
    Price from $9.99 to $1999.99
    hotstart taq dna polymerase - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    99
    Qiagen dna polymerase
    TALEN-induced mutations in the Platynereis estrogen receptor . (A) Schematic of the er locus showing target exons 2 and 3 with TALEN target sites (yellow). Blue arrows, primer positions; red double-ended arrow, region of sequence deleted in E; green, <t>DNA-binding</t> domain. Primer combinations used for screening are shown above in B–E. (B–E) <t>PCR,</t> undigested PCR product; NI, non-injected. (B) Restriction digest screening of larvae injected with er Ex3_L2/R2 TALENs (mRNA concentration: 267 ng/µl/TALEN mRNA). Arrowhead indicates uncut PCR product following AflI II digestion (asterisk). (C) Mutation evidence at exon 2 site: uncut band adult worm +3 vs. fully digested product from mutation-negative (−) TALEN-injected worm. (D) Adult worms er+31 and er+37 with mutations at exon 3 site. (E) Deletions (red arrow) detected in larvae and adult worms resulting from simultaneous cleavage at exons 2 and 3 using 300 ng/µl/TALEN mRNA: deletion positive (+); deletion negative (−). Please note different primer pairs used for larval vs. adult samples. (F) Mutant sequences obtained from digest screening for exons 1, 2, and long deletions. Numbers in brackets indicate the sample or worm from which the sequence was obtained; all other sequences are from injected larvae shown in B. Length of mutations are indicated by ∆ with “−” indicating deletions and “+” indicating insertions. Restriction site is shown in boldface type; asterisks indicate frameshift mutations. Shading key: yellow, TALEN binding sites; gray, spacer; blue, nucleotides differing from wild type; green, inserted nucleotides.
    Dna Polymerase, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 408 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna polymerase/product/Qiagen
    Average 99 stars, based on 408 article reviews
    Price from $9.99 to $1999.99
    dna polymerase - by Bioz Stars, 2020-04
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    Effect of individual constituents of the PCR on background fluorescence in the presence or absence of CABS. Each column represents the result of three PCRs performed: 1, in the absence of Taq Gold polymerase, but in the presence of oligonucleotides and template; 2, in the absence of Taq Gold polymerase and template, but in the presence of oligonucleotides; 3, in the absence of Taq Gold polymerase, template, and one oligonucleotide but in the presence of one oligonucleotide; 4, in the absence of primers and template, but in the presence of Taq Gold polymerase; 5, in the absence of template, but in the presence of both primers and polymerase; 6, as in case 3, except in the presence of the alternative primer of the primer combination; 7, in which specific amplicon is produced; and 8, in which no amplicon is expected. Conditions 1, 2, and 6 show the effect of CABS in reducing the background fluorescence caused by the single-strand oligonucleotides. The reaction with CABS in conditions 5 and 8 shows the residual effect caused by primer/dimers, in the case of 8 with additional template DNA.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Alkaline-mediated differential interaction (AMDI): A simple automatable single-nucleotide polymorphism assay

    doi: 10.1073/pnas.041619998

    Figure Lengend Snippet: Effect of individual constituents of the PCR on background fluorescence in the presence or absence of CABS. Each column represents the result of three PCRs performed: 1, in the absence of Taq Gold polymerase, but in the presence of oligonucleotides and template; 2, in the absence of Taq Gold polymerase and template, but in the presence of oligonucleotides; 3, in the absence of Taq Gold polymerase, template, and one oligonucleotide but in the presence of one oligonucleotide; 4, in the absence of primers and template, but in the presence of Taq Gold polymerase; 5, in the absence of template, but in the presence of both primers and polymerase; 6, as in case 3, except in the presence of the alternative primer of the primer combination; 7, in which specific amplicon is produced; and 8, in which no amplicon is expected. Conditions 1, 2, and 6 show the effect of CABS in reducing the background fluorescence caused by the single-strand oligonucleotides. The reaction with CABS in conditions 5 and 8 shows the residual effect caused by primer/dimers, in the case of 8 with additional template DNA.

    Article Snippet: The reaction mixture consisted of 67 mM Tris⋅HCl (pH 8.4) at 25°C, 16.6 mM ammonium sulfate, 2 mM magnesium chloride, 0.01% (vol/vol) Tween 20, 200 μM of each dNTP, 1 μM of each primer, and 0.5 units Hotstart Taq DNA polymerase (Qiagen) or AmpliTaq Gold (Perkin–Elmer).

    Techniques: Polymerase Chain Reaction, Fluorescence, Amplification, Produced

    TALEN-induced mutations in the Platynereis estrogen receptor . (A) Schematic of the er locus showing target exons 2 and 3 with TALEN target sites (yellow). Blue arrows, primer positions; red double-ended arrow, region of sequence deleted in E; green, DNA-binding domain. Primer combinations used for screening are shown above in B–E. (B–E) PCR, undigested PCR product; NI, non-injected. (B) Restriction digest screening of larvae injected with er Ex3_L2/R2 TALENs (mRNA concentration: 267 ng/µl/TALEN mRNA). Arrowhead indicates uncut PCR product following AflI II digestion (asterisk). (C) Mutation evidence at exon 2 site: uncut band adult worm +3 vs. fully digested product from mutation-negative (−) TALEN-injected worm. (D) Adult worms er+31 and er+37 with mutations at exon 3 site. (E) Deletions (red arrow) detected in larvae and adult worms resulting from simultaneous cleavage at exons 2 and 3 using 300 ng/µl/TALEN mRNA: deletion positive (+); deletion negative (−). Please note different primer pairs used for larval vs. adult samples. (F) Mutant sequences obtained from digest screening for exons 1, 2, and long deletions. Numbers in brackets indicate the sample or worm from which the sequence was obtained; all other sequences are from injected larvae shown in B. Length of mutations are indicated by ∆ with “−” indicating deletions and “+” indicating insertions. Restriction site is shown in boldface type; asterisks indicate frameshift mutations. Shading key: yellow, TALEN binding sites; gray, spacer; blue, nucleotides differing from wild type; green, inserted nucleotides.

    Journal: Genetics

    Article Title: TALENs Mediate Efficient and Heritable Mutation of Endogenous Genes in the Marine Annelid Platynereis dumerilii

    doi: 10.1534/genetics.113.161091

    Figure Lengend Snippet: TALEN-induced mutations in the Platynereis estrogen receptor . (A) Schematic of the er locus showing target exons 2 and 3 with TALEN target sites (yellow). Blue arrows, primer positions; red double-ended arrow, region of sequence deleted in E; green, DNA-binding domain. Primer combinations used for screening are shown above in B–E. (B–E) PCR, undigested PCR product; NI, non-injected. (B) Restriction digest screening of larvae injected with er Ex3_L2/R2 TALENs (mRNA concentration: 267 ng/µl/TALEN mRNA). Arrowhead indicates uncut PCR product following AflI II digestion (asterisk). (C) Mutation evidence at exon 2 site: uncut band adult worm +3 vs. fully digested product from mutation-negative (−) TALEN-injected worm. (D) Adult worms er+31 and er+37 with mutations at exon 3 site. (E) Deletions (red arrow) detected in larvae and adult worms resulting from simultaneous cleavage at exons 2 and 3 using 300 ng/µl/TALEN mRNA: deletion positive (+); deletion negative (−). Please note different primer pairs used for larval vs. adult samples. (F) Mutant sequences obtained from digest screening for exons 1, 2, and long deletions. Numbers in brackets indicate the sample or worm from which the sequence was obtained; all other sequences are from injected larvae shown in B. Length of mutations are indicated by ∆ with “−” indicating deletions and “+” indicating insertions. Restriction site is shown in boldface type; asterisks indicate frameshift mutations. Shading key: yellow, TALEN binding sites; gray, spacer; blue, nucleotides differing from wild type; green, inserted nucleotides.

    Article Snippet: PCR and restriction digest screening assays PCR reaction mixes contained DNA polymerase [either HotStar Taq Plus (Qiagen) or Phusion Polymerase (Fermentas)], 1.5–3 mM MgCl2 , 400 μM dNTPs, 200 μM of each primer, 1× reaction buffer (according to the enzyme used), 1–2 µl of DNA template in final volume of 25 or 50 µl.

    Techniques: Sequencing, Binding Assay, Polymerase Chain Reaction, Injection, TALENs, Concentration Assay, Mutagenesis

    Optimisation of the PPLMD method . The y-axis represents the average background-subtracted pixel density (0–64,000). A. Introduction of SpikeTarget DNA reduced background in negative control (purple vs. yellow). Blue: genomic DNA (containing RRS and MON810) + SpikeTarget, purple: MQ, yellow: MQ with SpikeTarget. All 4 padlock probes were present in the mix. HotStar Taq DNA polymerase was used to amplify in 40 LATE cycles. B. 80 LATE cycles increased signal compared to 40 cycles. Purple: 40 LATE cycles, blue: 80 LATE cycles. All 4 padlock probes were present in the mix (containing RRS and MON810). Vent ® exo - DNA polymerase was used.

    Journal: BMC Genomics

    Article Title: Optimised padlock probe ligation and microarray detection of multiple (non-authorised) GMOs in a single reaction

    doi: 10.1186/1471-2164-9-584

    Figure Lengend Snippet: Optimisation of the PPLMD method . The y-axis represents the average background-subtracted pixel density (0–64,000). A. Introduction of SpikeTarget DNA reduced background in negative control (purple vs. yellow). Blue: genomic DNA (containing RRS and MON810) + SpikeTarget, purple: MQ, yellow: MQ with SpikeTarget. All 4 padlock probes were present in the mix. HotStar Taq DNA polymerase was used to amplify in 40 LATE cycles. B. 80 LATE cycles increased signal compared to 40 cycles. Purple: 40 LATE cycles, blue: 80 LATE cycles. All 4 padlock probes were present in the mix (containing RRS and MON810). Vent ® exo - DNA polymerase was used.

    Article Snippet: The initial standard protocol with HotStar Taq DNA polymerase (Qiagen) as to amplify the circular padlock probe was altered by application of the 5'-3' exonuclease-deficient Vent® exo- DNA polymerase, theoretically allowing extended linear amplification of the circular padlock target molecule and subsequent logarithmic PCR amplification.

    Techniques: Negative Control

    DNase I hypersensitivity of the murine PWS-IC. DNase hypersensitive sites (DHS) were mapped by Southern blotting and indirect end-labeling after DNase I treatment of primary brain cells. The maternal and paternal alleles were analyzed separately using brain cells prepared from mice carrying a 35 kb PWS-IC deletion on either the paternal or maternal chromosome, respectively [8] , [18] . Cells were treated with increasing concentrations of DNase I; 0 DNase samples were purified genomic DNA from untreated brain cells. (A) Analysis by EcoRI digestion and hybridization with probe A. The thick arrows indicate the positions and sizes of prominent reproducible DNase I hypersensitive bands. Paternal indicates samples carrying a maternal PWS-IC deletion; Maternal indicates samples carrying a paternal PWS-C deletion. B) Analysis by Taq I digestion and hybridization with probe B. C) Analysis by Nco I digestion and hybridization with probe C. D) Summary of DNase I hypersensitive sites and their locations. The diagram depicts the Snrpn 5′ region showing relevant restriction enzyme sites and the positions of hybridization probes A, B and C (probe B is contained within probe C). The relative positions of DHS1-6 are indicated by short horizontal bars above the gene; all prominent DH sites (DHS1-DS6) are specific to the paternal chromosome. CAS denotes the conserved activator sequence [17] that co-localizes with DHS5. Thin horizontal lines below the gene depict regions deleted by targeted knockouts of the Snrpn locus [38] , [39] . The bent arrow depicts the location of the transcription initiation site. The position of each DH site relative to the transcription initiation site is estimated to be: DHS1, −3.2 kb; DHS2 includes the transcription initiation site; DHS3, +0.8 kb; DHS4, +1.4 kb; DHS5, +1.7 kb; DHS6, +4.1 kb; the positions of DHS1-DHS6 in the Snrpn gene were calculated as an average of at least two independent experiments and/or different restriction enzymes and probes. The same DH sites mapped by different experiments, restriction enzymes, and/or blots generally localized within 200–300 bp of each other, a variation within the range expected by estimating positions derived from band sizes in Southern blots.

    Journal: PLoS ONE

    Article Title: Regulatory Elements Associated with Paternally-Expressed Genes in the Imprinted Murine Angelman/Prader-Willi Syndrome Domain

    doi: 10.1371/journal.pone.0052390

    Figure Lengend Snippet: DNase I hypersensitivity of the murine PWS-IC. DNase hypersensitive sites (DHS) were mapped by Southern blotting and indirect end-labeling after DNase I treatment of primary brain cells. The maternal and paternal alleles were analyzed separately using brain cells prepared from mice carrying a 35 kb PWS-IC deletion on either the paternal or maternal chromosome, respectively [8] , [18] . Cells were treated with increasing concentrations of DNase I; 0 DNase samples were purified genomic DNA from untreated brain cells. (A) Analysis by EcoRI digestion and hybridization with probe A. The thick arrows indicate the positions and sizes of prominent reproducible DNase I hypersensitive bands. Paternal indicates samples carrying a maternal PWS-IC deletion; Maternal indicates samples carrying a paternal PWS-C deletion. B) Analysis by Taq I digestion and hybridization with probe B. C) Analysis by Nco I digestion and hybridization with probe C. D) Summary of DNase I hypersensitive sites and their locations. The diagram depicts the Snrpn 5′ region showing relevant restriction enzyme sites and the positions of hybridization probes A, B and C (probe B is contained within probe C). The relative positions of DHS1-6 are indicated by short horizontal bars above the gene; all prominent DH sites (DHS1-DS6) are specific to the paternal chromosome. CAS denotes the conserved activator sequence [17] that co-localizes with DHS5. Thin horizontal lines below the gene depict regions deleted by targeted knockouts of the Snrpn locus [38] , [39] . The bent arrow depicts the location of the transcription initiation site. The position of each DH site relative to the transcription initiation site is estimated to be: DHS1, −3.2 kb; DHS2 includes the transcription initiation site; DHS3, +0.8 kb; DHS4, +1.4 kb; DHS5, +1.7 kb; DHS6, +4.1 kb; the positions of DHS1-DHS6 in the Snrpn gene were calculated as an average of at least two independent experiments and/or different restriction enzymes and probes. The same DH sites mapped by different experiments, restriction enzymes, and/or blots generally localized within 200–300 bp of each other, a variation within the range expected by estimating positions derived from band sizes in Southern blots.

    Article Snippet: For a 25 ul PCR reaction using 3C primers, 1–2 µl template DNA was mixed with 10 pmol of each primer, 5 nmol of each dATP, dTTP, dGTP, dCTP, 0.625 units of Hotstar Taq DNA polymerase (Qiagen).

    Techniques: Southern Blot, End Labeling, Mouse Assay, Purification, Hybridization, Sequencing, Derivative Assay