hotstart taq  (Qiagen)


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    Structured Review

    Qiagen hotstart taq
    Hotstart Taq, supplied by Qiagen, used in various techniques. Bioz Stars score: 96/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hotstart taq/product/Qiagen
    Average 96 stars, based on 54 article reviews
    Price from $9.99 to $1999.99
    hotstart taq - by Bioz Stars, 2020-04
    96/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data
    Article Snippet: Paragraph title: Purification, cloning and BS sequencing of the major satellite repeat ... The major satellite was amplified with HotStart Taq (Qiagen) in a mixture of 200 nM primer, 200 μM dNTPs, 2 mM MgCl2 , 1.0 unit of enzyme at 94 °C for 15 min, 35 cycles of 20 s at 94 °C, 20 s at 55 °C, and 20 s at 72 °C, and a final step at 72 °C for 3 min. DNA fragments spanning over one repeat (370 bp) were excised from 2% agarose gels and purified with a MinElute Gel Extraction kit (Qiagen) following the kit protocol.

    Amplification:

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: .. Followed by Hotstart Taq (Qiagen) activation at 95°C for 15 min, the first round of PCR was performed for 50 cycles of 94°C for 30s, 56°C for 30s, 72°C for 55s, and final extension at 72°C for 10 min. Primers for amplification were 5′ MsVHE (5′-GGGAATTCGAGGTGCAGCTGCAGGAGTCTGG-3′) and 3′ Cμ outer. .. 2 µl of unpurified first round PCR product was used for the seminested second round PCR, starting with Hotstart Taq (Qiagen) activation at 95°C for 15 min.

    Article Title: Carriers of the Complex Allele HFE c.[187C > G;340+4T > C] Have Increased Risk of Iron Overload in São Miguel Island Population (Azores, Portugal)
    Article Snippet: .. Each 20 μl PCR amplification reaction contained 100 ng of genomic DNA, 10 μM primers, 200 μM dNTPs (Promega), 25 nM MgCl2 (Qiagen), 1x Q-Solution (Qiagen), 1x buffer (Qiagen), 5 U of HotStart Taq (Qiagen), and sterile water. .. The PCR started with an enzyme activation step at 95°C for 15 min, then 40 cycles at 94°C for 30s, 56.5°C for 30s and 72°C for 1 min, followed by a final extension step at 72°C for 10 min. Amplification products were purified using the QIAquick PCR Purification kit (Qiagen), according to the manufacturer’s instructions.

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: Paragraph title: RT-PCR amplification of VH genes from single cell culture ... 2 µl of unpurified first round PCR product was used for the seminested second round PCR, starting with Hotstart Taq (Qiagen) activation at 95°C for 15 min.

    Article Title: RFLP Typing Demonstrates Substantial Diversity among Pneumocystis jirovecii Isolates
    Article Snippet: Paragraph title: DNA amplification of msg variable region ... The PCR was performed using HotStart Taq (Qiagen) and the conditions were 15 min at 95°C followed by 35 cycles of 30 s at 94°C, 30 s at 60°C, and 4 min (for the first round) or 2 min (for the second round) at 72°C, with a finally extension of 10 min at 72°C. msg copy number was quantitated by a previously described real-time Q-PCR assay [ ].

    Article Title: Evidence that the Essential Response Regulator YycF in Streptococcus pneumoniae Modulates Expression of Fatty Acid Biosynthesis Genes and Alters Membrane Composition †
    Article Snippet: In total, 2,236 TIGR4 and 117 R6 gene-specific probes were amplified by PCR for spotting onto the microarray. .. All PCRs were performed with HotStart Taq (QIAGEN) and a Primus-HT PCR machine (MWG Biotech) according to the following parameters: 95°C for 15 min, followed by 30 cycles of 95°C for 30 s, 50°C for 30 s, and 72°C for 120 s, with a final incubation at 72°C for 600 s. Genomic TIGR4 and R6 DNAs (2.5 ng) were used as templates, and approximately 100 pmol of each primer was used in a reaction volume of 50 μl.

    Article Title: Anaplastic large cell lymphoma arises in thymocytes and requires transient TCR expression for thymic egress
    Article Snippet: .. RNA (20 ng) was amplified in each PCR with 1 U of Hotstart Taq (Qiagen, Courtaboeuf, France), 10 mM dNTPs, 2 mM of MgCl2 , 20% Q solution, and 10 pmol of each primer. ..

    Article Title: Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data
    Article Snippet: .. The major satellite was amplified with HotStart Taq (Qiagen) in a mixture of 200 nM primer, 200 μM dNTPs, 2 mM MgCl2 , 1.0 unit of enzyme at 94 °C for 15 min, 35 cycles of 20 s at 94 °C, 20 s at 55 °C, and 20 s at 72 °C, and a final step at 72 °C for 3 min. DNA fragments spanning over one repeat (370 bp) were excised from 2% agarose gels and purified with a MinElute Gel Extraction kit (Qiagen) following the kit protocol. .. The fragments were cloned into pGEM-T using the pGEM-T Easy Vector Kit (Promega) and transformed into Invitrogen’s Subcloning Efficiency DH5α Competent Cells according to the manufacturer’s instructions.

    Article Title: Diagnosis of Human Leptospirosis in a Clinical Setting: Real-Time PCR High Resolution Melting Analysis for Detection of Leptospira at the Onset of Disease
    Article Snippet: After automatic bacterial DNA extraction, the rrs (16S rRNA) gene was amplified as previously described , by conventional nested PCR in a Biometra® T-Gradient thermal cycler. .. The first PCR reaction contained 5 μl of bacterial DNA, 10 μM primers A and B, 100 μM dNTPs (Promega), 25 nM MgCl2 (Qiagen), 1X Q-Solution (Qiagen), 1X buffer (Qiagen), 5 U of HotStart Taq (Qiagen) and RNase-free water to a final volume of 50 μl.

    Article Title: Streptococcus pneumoniae induced c-Jun-N-terminal kinase- and AP-1 -dependent IL-8 release by lung epithelial BEAS-2B cells
    Article Snippet: .. After proteinase K digestion (1 μg/8 μl for 6 h at 37°C and 6 h at 65°) DNA was extracted using a PCR purification kit (Qiagen, Hilden, Germany). il8 promoter DNA was amplified by PCR using Hotstart Taq (Qiagen) polymerase. .. The PCR conditions were 95°C for 15 min, 33 – 35 cycles of 94°C for 20 s, 60°C for 20 s, 72°C for 20 s. PCR products were separated by agarose gel electrophoresis and detected by ethidium bromide staining.

    Positive Control:

    Article Title: Anaplastic large cell lymphoma arises in thymocytes and requires transient TCR expression for thymic egress
    Article Snippet: RNA (20 ng) was amplified in each PCR with 1 U of Hotstart Taq (Qiagen, Courtaboeuf, France), 10 mM dNTPs, 2 mM of MgCl2 , 20% Q solution, and 10 pmol of each primer. .. The negative control consisted of RNA isolated from peripheral blood mononuclear cells from donors and the positive control was RNA isolated from the Jurkat cell line.

    Synthesized:

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: The first-strand cDNA was synthesized in a final volume of 10 µl/well using 10 µM primer specific for the μ constant region (3′ Cμ outer, 5′-AGGGGGCTCTCGCAGGAGACGAGG-3′; ),1 mM dNTP (Promega), 10 mM dithiothreitol (Thermo Fisher), 0.5% vol/vol Igepal CA-630 (Sigma), 12 U RNAaseout (Thermo Fisher), and 40 U Superscript III reverse transcription (Thermo Fisher). .. Followed by Hotstart Taq (Qiagen) activation at 95°C for 15 min, the first round of PCR was performed for 50 cycles of 94°C for 30s, 56°C for 30s, 72°C for 55s, and final extension at 72°C for 10 min. Primers for amplification were 5′ MsVHE (5′-GGGAATTCGAGGTGCAGCTGCAGGAGTCTGG-3′) and 3′ Cμ outer.

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: The first-strand cDNA was synthesized in a final volume of 10 µl/well using 10 µM primer specific for the μ constant region (3′ Cμ outer, 5′-AGGGGGCTCTCGCAGGAGACGAGG-3′; ),1 mM dNTP (Promega), 10 mM dithiothreitol (Thermo Fisher), 0.5% vol/vol Igepal CA-630 (Sigma), 12 U RNAaseout (Thermo Fisher), and 40 U Superscript III reverse transcription (Thermo Fisher). .. 2 µl of unpurified first round PCR product was used for the seminested second round PCR, starting with Hotstart Taq (Qiagen) activation at 95°C for 15 min.

    Electrophoresis:

    Article Title: Evidence that the Essential Response Regulator YycF in Streptococcus pneumoniae Modulates Expression of Fatty Acid Biosynthesis Genes and Alters Membrane Composition †
    Article Snippet: All PCRs were performed with HotStart Taq (QIAGEN) and a Primus-HT PCR machine (MWG Biotech) according to the following parameters: 95°C for 15 min, followed by 30 cycles of 95°C for 30 s, 50°C for 30 s, and 72°C for 120 s, with a final incubation at 72°C for 600 s. Genomic TIGR4 and R6 DNAs (2.5 ng) were used as templates, and approximately 100 pmol of each primer was used in a reaction volume of 50 μl. .. For each primer pair, the successful PCR amplification of a single DNA fragment of the correct length was checked by electrophoresis in agarose gels.

    Microarray:

    Article Title: Evidence that the Essential Response Regulator YycF in Streptococcus pneumoniae Modulates Expression of Fatty Acid Biosynthesis Genes and Alters Membrane Composition †
    Article Snippet: Paragraph title: Construction of S. pneumoniae microarray. ... All PCRs were performed with HotStart Taq (QIAGEN) and a Primus-HT PCR machine (MWG Biotech) according to the following parameters: 95°C for 15 min, followed by 30 cycles of 95°C for 30 s, 50°C for 30 s, and 72°C for 120 s, with a final incubation at 72°C for 600 s. Genomic TIGR4 and R6 DNAs (2.5 ng) were used as templates, and approximately 100 pmol of each primer was used in a reaction volume of 50 μl.

    Incubation:

    Article Title: Evidence that the Essential Response Regulator YycF in Streptococcus pneumoniae Modulates Expression of Fatty Acid Biosynthesis Genes and Alters Membrane Composition †
    Article Snippet: .. All PCRs were performed with HotStart Taq (QIAGEN) and a Primus-HT PCR machine (MWG Biotech) according to the following parameters: 95°C for 15 min, followed by 30 cycles of 95°C for 30 s, 50°C for 30 s, and 72°C for 120 s, with a final incubation at 72°C for 600 s. Genomic TIGR4 and R6 DNAs (2.5 ng) were used as templates, and approximately 100 pmol of each primer was used in a reaction volume of 50 μl. .. For each primer pair, the successful PCR amplification of a single DNA fragment of the correct length was checked by electrophoresis in agarose gels.

    Activity Assay:

    Article Title: Cold-sensitive mutants of Taq DNA polymerase provide a hot start for PCR
    Article Snippet: The antibodies available for this method are not very efficient and must therefore be used in a 5- or 10-fold molar excess or in a triple combination to effectively reduce the activity of the DNA polymerase. .. Some enzyme preparations, such as Amplitaq Gold (Perkin Elmer and Roche Molecular Systems) and Hotstart Taq (Qiagen) are chemically inactivated and can be reactivated by heating in a special preincubation step ( ).

    Transformation Assay:

    Article Title: Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data
    Article Snippet: The major satellite was amplified with HotStart Taq (Qiagen) in a mixture of 200 nM primer, 200 μM dNTPs, 2 mM MgCl2 , 1.0 unit of enzyme at 94 °C for 15 min, 35 cycles of 20 s at 94 °C, 20 s at 55 °C, and 20 s at 72 °C, and a final step at 72 °C for 3 min. DNA fragments spanning over one repeat (370 bp) were excised from 2% agarose gels and purified with a MinElute Gel Extraction kit (Qiagen) following the kit protocol. .. The fragments were cloned into pGEM-T using the pGEM-T Easy Vector Kit (Promega) and transformed into Invitrogen’s Subcloning Efficiency DH5α Competent Cells according to the manufacturer’s instructions.

    Countercurrent Chromatography:

    Article Title: Streptococcus pneumoniae induced c-Jun-N-terminal kinase- and AP-1 -dependent IL-8 release by lung epithelial BEAS-2B cells
    Article Snippet: After proteinase K digestion (1 μg/8 μl for 6 h at 37°C and 6 h at 65°) DNA was extracted using a PCR purification kit (Qiagen, Hilden, Germany). il8 promoter DNA was amplified by PCR using Hotstart Taq (Qiagen) polymerase. .. The following il8 promoter-specific primers were used: sense 5'-AAG AAA ACT TTC GTC ATA CTC CG-3'; antisense 5'-TGG CTT TTT ATA TCA TCA CCC TAC-3' [ ].

    Transferring:

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: After transferring the supernatants, the pellets were lysed with 10 µl/well of ice-cold 0.5 × PBS containing 10mM dithiothreitol and 4 U RNAaseout (Thermo Fisher). .. Followed by Hotstart Taq (Qiagen) activation at 95°C for 15 min, the first round of PCR was performed for 50 cycles of 94°C for 30s, 56°C for 30s, 72°C for 55s, and final extension at 72°C for 10 min. Primers for amplification were 5′ MsVHE (5′-GGGAATTCGAGGTGCAGCTGCAGGAGTCTGG-3′) and 3′ Cμ outer.

    Cell Culture:

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: Paragraph title: RT-PCR amplification of VH genes from single cell culture ... Followed by Hotstart Taq (Qiagen) activation at 95°C for 15 min, the first round of PCR was performed for 50 cycles of 94°C for 30s, 56°C for 30s, 72°C for 55s, and final extension at 72°C for 10 min. Primers for amplification were 5′ MsVHE (5′-GGGAATTCGAGGTGCAGCTGCAGGAGTCTGG-3′) and 3′ Cμ outer.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: Paragraph title: RT-PCR amplification of VH genes from single cell culture ... Followed by Hotstart Taq (Qiagen) activation at 95°C for 15 min, the first round of PCR was performed for 50 cycles of 94°C for 30s, 56°C for 30s, 72°C for 55s, and final extension at 72°C for 10 min. Primers for amplification were 5′ MsVHE (5′-GGGAATTCGAGGTGCAGCTGCAGGAGTCTGG-3′) and 3′ Cμ outer.

    Article Title: Anaplastic large cell lymphoma arises in thymocytes and requires transient TCR expression for thymic egress
    Article Snippet: TCRα rearrangements were amplified from cDNA using Cα and Vα primers in five multiplex RT–PCR reactions . .. RNA (20 ng) was amplified in each PCR with 1 U of Hotstart Taq (Qiagen, Courtaboeuf, France), 10 mM dNTPs, 2 mM of MgCl2 , 20% Q solution, and 10 pmol of each primer.

    Article Title: Two TonB Systems in Actinobacillus pleuropneumoniae: Their Roles in Iron Acquisition and Virulence
    Article Snippet: .. Negative control experiments to test for the presence of contaminating DNA were performed by incubating the RT-PCR mixture (prior to the addition of enzyme) with 4 μl of RNase A (100 mg/ml) for 15 min at 37°C or by using HotStart Taq (Qiagen) instead of the supplied enzyme mixture, thus eliminating the reverse transcriptase step. .. tonB1 was identified in a cosmid library of A. pleuropneumoniae serotype 1 by using a probe based on the published partial sequence , and tonB1 and associated genes were sequenced directly from the clone.

    Sequencing:

    Article Title: Carriers of the Complex Allele HFE c.[187C > G;340+4T > C] Have Increased Risk of Iron Overload in São Miguel Island Population (Azores, Portugal)
    Article Snippet: The exon 2 and its intron-flanking sequences analysis allows for the identification of the c.340+4T > C [IVS2+4T > C] splice site variation, which has been reported as associated with iron overload [ ], whereas sequencing of exon 5 and its intron-flanking regions allows for the evaluation of the splice site mutation c.1008+1G > A [IVS5+1G > A] [ ]. .. Each 20 μl PCR amplification reaction contained 100 ng of genomic DNA, 10 μM primers, 200 μM dNTPs (Promega), 25 nM MgCl2 (Qiagen), 1x Q-Solution (Qiagen), 1x buffer (Qiagen), 5 U of HotStart Taq (Qiagen), and sterile water.

    Article Title: Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data
    Article Snippet: Paragraph title: Purification, cloning and BS sequencing of the major satellite repeat ... The major satellite was amplified with HotStart Taq (Qiagen) in a mixture of 200 nM primer, 200 μM dNTPs, 2 mM MgCl2 , 1.0 unit of enzyme at 94 °C for 15 min, 35 cycles of 20 s at 94 °C, 20 s at 55 °C, and 20 s at 72 °C, and a final step at 72 °C for 3 min. DNA fragments spanning over one repeat (370 bp) were excised from 2% agarose gels and purified with a MinElute Gel Extraction kit (Qiagen) following the kit protocol.

    DNA Extraction:

    Article Title: Diagnosis of Human Leptospirosis in a Clinical Setting: Real-Time PCR High Resolution Melting Analysis for Detection of Leptospira at the Onset of Disease
    Article Snippet: After automatic bacterial DNA extraction, the rrs (16S rRNA) gene was amplified as previously described , by conventional nested PCR in a Biometra® T-Gradient thermal cycler. .. The first PCR reaction contained 5 μl of bacterial DNA, 10 μM primers A and B, 100 μM dNTPs (Promega), 25 nM MgCl2 (Qiagen), 1X Q-Solution (Qiagen), 1X buffer (Qiagen), 5 U of HotStart Taq (Qiagen) and RNase-free water to a final volume of 50 μl.

    Nucleic Acid Electrophoresis:

    Article Title: Streptococcus pneumoniae induced c-Jun-N-terminal kinase- and AP-1 -dependent IL-8 release by lung epithelial BEAS-2B cells
    Article Snippet: After proteinase K digestion (1 μg/8 μl for 6 h at 37°C and 6 h at 65°) DNA was extracted using a PCR purification kit (Qiagen, Hilden, Germany). il8 promoter DNA was amplified by PCR using Hotstart Taq (Qiagen) polymerase. .. Equal amounts of input DNA was controlled by gel electrophoresis.

    Mutagenesis:

    Article Title: Carriers of the Complex Allele HFE c.[187C > G;340+4T > C] Have Increased Risk of Iron Overload in São Miguel Island Population (Azores, Portugal)
    Article Snippet: Paragraph title: HFE mutation analysis ... Each 20 μl PCR amplification reaction contained 100 ng of genomic DNA, 10 μM primers, 200 μM dNTPs (Promega), 25 nM MgCl2 (Qiagen), 1x Q-Solution (Qiagen), 1x buffer (Qiagen), 5 U of HotStart Taq (Qiagen), and sterile water.

    Isolation:

    Article Title: Anaplastic large cell lymphoma arises in thymocytes and requires transient TCR expression for thymic egress
    Article Snippet: RNA was isolated using an RNeasy kit according to the manufacturer's instructions (Qiagen, Courtaboeuf, France) and 1 μg of total RNA was converted into cDNA using SuperScript III RT (Invitrogen, San Diego, USA). .. RNA (20 ng) was amplified in each PCR with 1 U of Hotstart Taq (Qiagen, Courtaboeuf, France), 10 mM dNTPs, 2 mM of MgCl2 , 20% Q solution, and 10 pmol of each primer.

    Subcloning:

    Article Title: Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data
    Article Snippet: The major satellite was amplified with HotStart Taq (Qiagen) in a mixture of 200 nM primer, 200 μM dNTPs, 2 mM MgCl2 , 1.0 unit of enzyme at 94 °C for 15 min, 35 cycles of 20 s at 94 °C, 20 s at 55 °C, and 20 s at 72 °C, and a final step at 72 °C for 3 min. DNA fragments spanning over one repeat (370 bp) were excised from 2% agarose gels and purified with a MinElute Gel Extraction kit (Qiagen) following the kit protocol. .. The fragments were cloned into pGEM-T using the pGEM-T Easy Vector Kit (Promega) and transformed into Invitrogen’s Subcloning Efficiency DH5α Competent Cells according to the manufacturer’s instructions.

    Multiplex Assay:

    Article Title: Anaplastic large cell lymphoma arises in thymocytes and requires transient TCR expression for thymic egress
    Article Snippet: TCRα rearrangements were amplified from cDNA using Cα and Vα primers in five multiplex RT–PCR reactions . .. RNA (20 ng) was amplified in each PCR with 1 U of Hotstart Taq (Qiagen, Courtaboeuf, France), 10 mM dNTPs, 2 mM of MgCl2 , 20% Q solution, and 10 pmol of each primer.

    Purification:

    Article Title: Carriers of the Complex Allele HFE c.[187C > G;340+4T > C] Have Increased Risk of Iron Overload in São Miguel Island Population (Azores, Portugal)
    Article Snippet: Each 20 μl PCR amplification reaction contained 100 ng of genomic DNA, 10 μM primers, 200 μM dNTPs (Promega), 25 nM MgCl2 (Qiagen), 1x Q-Solution (Qiagen), 1x buffer (Qiagen), 5 U of HotStart Taq (Qiagen), and sterile water. .. The PCR started with an enzyme activation step at 95°C for 15 min, then 40 cycles at 94°C for 30s, 56.5°C for 30s and 72°C for 1 min, followed by a final extension step at 72°C for 10 min. Amplification products were purified using the QIAquick PCR Purification kit (Qiagen), according to the manufacturer’s instructions.

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: 2 µl of unpurified first round PCR product was used for the seminested second round PCR, starting with Hotstart Taq (Qiagen) activation at 95°C for 15 min. .. Aliquots of the PCR product were purified using QIAquick PCR purification kit (Qiagen) and sequenced with the reverse primer 3′ Cμ inner.

    Article Title: Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data
    Article Snippet: .. The major satellite was amplified with HotStart Taq (Qiagen) in a mixture of 200 nM primer, 200 μM dNTPs, 2 mM MgCl2 , 1.0 unit of enzyme at 94 °C for 15 min, 35 cycles of 20 s at 94 °C, 20 s at 55 °C, and 20 s at 72 °C, and a final step at 72 °C for 3 min. DNA fragments spanning over one repeat (370 bp) were excised from 2% agarose gels and purified with a MinElute Gel Extraction kit (Qiagen) following the kit protocol. .. The fragments were cloned into pGEM-T using the pGEM-T Easy Vector Kit (Promega) and transformed into Invitrogen’s Subcloning Efficiency DH5α Competent Cells according to the manufacturer’s instructions.

    Article Title: Streptococcus pneumoniae induced c-Jun-N-terminal kinase- and AP-1 -dependent IL-8 release by lung epithelial BEAS-2B cells
    Article Snippet: .. After proteinase K digestion (1 μg/8 μl for 6 h at 37°C and 6 h at 65°) DNA was extracted using a PCR purification kit (Qiagen, Hilden, Germany). il8 promoter DNA was amplified by PCR using Hotstart Taq (Qiagen) polymerase. .. The PCR conditions were 95°C for 15 min, 33 – 35 cycles of 94°C for 20 s, 60°C for 20 s, 72°C for 20 s. PCR products were separated by agarose gel electrophoresis and detected by ethidium bromide staining.

    Polymerase Chain Reaction:

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: .. Followed by Hotstart Taq (Qiagen) activation at 95°C for 15 min, the first round of PCR was performed for 50 cycles of 94°C for 30s, 56°C for 30s, 72°C for 55s, and final extension at 72°C for 10 min. Primers for amplification were 5′ MsVHE (5′-GGGAATTCGAGGTGCAGCTGCAGGAGTCTGG-3′) and 3′ Cμ outer. .. 2 µl of unpurified first round PCR product was used for the seminested second round PCR, starting with Hotstart Taq (Qiagen) activation at 95°C for 15 min.

    Article Title: Carriers of the Complex Allele HFE c.[187C > G;340+4T > C] Have Increased Risk of Iron Overload in São Miguel Island Population (Azores, Portugal)
    Article Snippet: .. Each 20 μl PCR amplification reaction contained 100 ng of genomic DNA, 10 μM primers, 200 μM dNTPs (Promega), 25 nM MgCl2 (Qiagen), 1x Q-Solution (Qiagen), 1x buffer (Qiagen), 5 U of HotStart Taq (Qiagen), and sterile water. .. The PCR started with an enzyme activation step at 95°C for 15 min, then 40 cycles at 94°C for 30s, 56.5°C for 30s and 72°C for 1 min, followed by a final extension step at 72°C for 10 min. Amplification products were purified using the QIAquick PCR Purification kit (Qiagen), according to the manufacturer’s instructions.

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: .. 2 µl of unpurified first round PCR product was used for the seminested second round PCR, starting with Hotstart Taq (Qiagen) activation at 95°C for 15 min. .. The reactions were then performed for 50 cycles of 94°C for 30 s, 45°C for 30 s, 72°C for 45 s, and final extension at 72°C for 10 min. Primers for amplification were 5′ MsVHE and 3′ Cμ inner (5′-AGGGGGAAGACATTTGGGAAGGAC-3′; ).

    Article Title: Cold-sensitive mutants of Taq DNA polymerase provide a hot start for PCR
    Article Snippet: Anti-Taq antibodies reduce the DNA polymerase activity but, being thermolabile, release the enzyme at PCR cycling temperatures to achieve a hot start ( – ). .. Some enzyme preparations, such as Amplitaq Gold (Perkin Elmer and Roche Molecular Systems) and Hotstart Taq (Qiagen) are chemically inactivated and can be reactivated by heating in a special preincubation step ( ).

    Article Title: RFLP Typing Demonstrates Substantial Diversity among Pneumocystis jirovecii Isolates
    Article Snippet: .. The PCR was performed using HotStart Taq (Qiagen) and the conditions were 15 min at 95°C followed by 35 cycles of 30 s at 94°C, 30 s at 60°C, and 4 min (for the first round) or 2 min (for the second round) at 72°C, with a finally extension of 10 min at 72°C. msg copy number was quantitated by a previously described real-time Q-PCR assay [ ]. .. PCR products were purified using QuickStep 2 PCR Purification Kit (Edge BioSystems, Gaithersburg, Maryland) according to the manufacturer's instructions.

    Article Title: Evidence that the Essential Response Regulator YycF in Streptococcus pneumoniae Modulates Expression of Fatty Acid Biosynthesis Genes and Alters Membrane Composition †
    Article Snippet: .. All PCRs were performed with HotStart Taq (QIAGEN) and a Primus-HT PCR machine (MWG Biotech) according to the following parameters: 95°C for 15 min, followed by 30 cycles of 95°C for 30 s, 50°C for 30 s, and 72°C for 120 s, with a final incubation at 72°C for 600 s. Genomic TIGR4 and R6 DNAs (2.5 ng) were used as templates, and approximately 100 pmol of each primer was used in a reaction volume of 50 μl. .. For each primer pair, the successful PCR amplification of a single DNA fragment of the correct length was checked by electrophoresis in agarose gels.

    Article Title: Anaplastic large cell lymphoma arises in thymocytes and requires transient TCR expression for thymic egress
    Article Snippet: .. RNA (20 ng) was amplified in each PCR with 1 U of Hotstart Taq (Qiagen, Courtaboeuf, France), 10 mM dNTPs, 2 mM of MgCl2 , 20% Q solution, and 10 pmol of each primer. ..

    Article Title: Diagnosis of Human Leptospirosis in a Clinical Setting: Real-Time PCR High Resolution Melting Analysis for Detection of Leptospira at the Onset of Disease
    Article Snippet: .. The first PCR reaction contained 5 μl of bacterial DNA, 10 μM primers A and B, 100 μM dNTPs (Promega), 25 nM MgCl2 (Qiagen), 1X Q-Solution (Qiagen), 1X buffer (Qiagen), 5 U of HotStart Taq (Qiagen) and RNase-free water to a final volume of 50 μl. .. The PCR programme started with an enzyme activation step at 95 °C for 15 minutes; proceeded with 30 cycles of 94 °C for 1 minute, 63 °C for 1 minute and 72 °C for 1 minute; and ended with a final extension step at 72 °C for 10 minutes.

    Article Title: Two TonB Systems in Actinobacillus pleuropneumoniae: Their Roles in Iron Acquisition and Virulence
    Article Snippet: Paragraph title: PCR and RT-PCR. ... Negative control experiments to test for the presence of contaminating DNA were performed by incubating the RT-PCR mixture (prior to the addition of enzyme) with 4 μl of RNase A (100 mg/ml) for 15 min at 37°C or by using HotStart Taq (Qiagen) instead of the supplied enzyme mixture, thus eliminating the reverse transcriptase step.

    Article Title: Streptococcus pneumoniae induced c-Jun-N-terminal kinase- and AP-1 -dependent IL-8 release by lung epithelial BEAS-2B cells
    Article Snippet: .. After proteinase K digestion (1 μg/8 μl for 6 h at 37°C and 6 h at 65°) DNA was extracted using a PCR purification kit (Qiagen, Hilden, Germany). il8 promoter DNA was amplified by PCR using Hotstart Taq (Qiagen) polymerase. .. The PCR conditions were 95°C for 15 min, 33 – 35 cycles of 94°C for 20 s, 60°C for 20 s, 72°C for 20 s. PCR products were separated by agarose gel electrophoresis and detected by ethidium bromide staining.

    Gel Extraction:

    Article Title: Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data
    Article Snippet: .. The major satellite was amplified with HotStart Taq (Qiagen) in a mixture of 200 nM primer, 200 μM dNTPs, 2 mM MgCl2 , 1.0 unit of enzyme at 94 °C for 15 min, 35 cycles of 20 s at 94 °C, 20 s at 55 °C, and 20 s at 72 °C, and a final step at 72 °C for 3 min. DNA fragments spanning over one repeat (370 bp) were excised from 2% agarose gels and purified with a MinElute Gel Extraction kit (Qiagen) following the kit protocol. .. The fragments were cloned into pGEM-T using the pGEM-T Easy Vector Kit (Promega) and transformed into Invitrogen’s Subcloning Efficiency DH5α Competent Cells according to the manufacturer’s instructions.

    Nested PCR:

    Article Title: Diagnosis of Human Leptospirosis in a Clinical Setting: Real-Time PCR High Resolution Melting Analysis for Detection of Leptospira at the Onset of Disease
    Article Snippet: Paragraph title: Reference molecular test (conventional nested PCR) ... The first PCR reaction contained 5 μl of bacterial DNA, 10 μM primers A and B, 100 μM dNTPs (Promega), 25 nM MgCl2 (Qiagen), 1X Q-Solution (Qiagen), 1X buffer (Qiagen), 5 U of HotStart Taq (Qiagen) and RNase-free water to a final volume of 50 μl.

    Activated Clotting Time Assay:

    Article Title: Streptococcus pneumoniae induced c-Jun-N-terminal kinase- and AP-1 -dependent IL-8 release by lung epithelial BEAS-2B cells
    Article Snippet: After proteinase K digestion (1 μg/8 μl for 6 h at 37°C and 6 h at 65°) DNA was extracted using a PCR purification kit (Qiagen, Hilden, Germany). il8 promoter DNA was amplified by PCR using Hotstart Taq (Qiagen) polymerase. .. The following il8 promoter-specific primers were used: sense 5'-AAG AAA ACT TTC GTC ATA CTC CG-3'; antisense 5'-TGG CTT TTT ATA TCA TCA CCC TAC-3' [ ].

    Chromatin Immunoprecipitation:

    Article Title: Streptococcus pneumoniae induced c-Jun-N-terminal kinase- and AP-1 -dependent IL-8 release by lung epithelial BEAS-2B cells
    Article Snippet: Paragraph title: Chromatin immunoprecipitation ... After proteinase K digestion (1 μg/8 μl for 6 h at 37°C and 6 h at 65°) DNA was extracted using a PCR purification kit (Qiagen, Hilden, Germany). il8 promoter DNA was amplified by PCR using Hotstart Taq (Qiagen) polymerase.

    Plasmid Preparation:

    Article Title: Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data
    Article Snippet: The major satellite was amplified with HotStart Taq (Qiagen) in a mixture of 200 nM primer, 200 μM dNTPs, 2 mM MgCl2 , 1.0 unit of enzyme at 94 °C for 15 min, 35 cycles of 20 s at 94 °C, 20 s at 55 °C, and 20 s at 72 °C, and a final step at 72 °C for 3 min. DNA fragments spanning over one repeat (370 bp) were excised from 2% agarose gels and purified with a MinElute Gel Extraction kit (Qiagen) following the kit protocol. .. The fragments were cloned into pGEM-T using the pGEM-T Easy Vector Kit (Promega) and transformed into Invitrogen’s Subcloning Efficiency DH5α Competent Cells according to the manufacturer’s instructions.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: Based on the ELISA results, 4 µl of the lysates from all the original wells producing bacteria-reactive IgM and a proportion of the wells producing bacteria-nonreactive IgM were processed for reverse transcription. .. Followed by Hotstart Taq (Qiagen) activation at 95°C for 15 min, the first round of PCR was performed for 50 cycles of 94°C for 30s, 56°C for 30s, 72°C for 55s, and final extension at 72°C for 10 min. Primers for amplification were 5′ MsVHE (5′-GGGAATTCGAGGTGCAGCTGCAGGAGTCTGG-3′) and 3′ Cμ outer.

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: Based on the ELISA results, 4 µl of the lysates from all the original wells producing bacteria-reactive IgM and a proportion of the wells producing bacteria-nonreactive IgM were processed for reverse transcription. .. 2 µl of unpurified first round PCR product was used for the seminested second round PCR, starting with Hotstart Taq (Qiagen) activation at 95°C for 15 min.

    Negative Control:

    Article Title: Anaplastic large cell lymphoma arises in thymocytes and requires transient TCR expression for thymic egress
    Article Snippet: RNA (20 ng) was amplified in each PCR with 1 U of Hotstart Taq (Qiagen, Courtaboeuf, France), 10 mM dNTPs, 2 mM of MgCl2 , 20% Q solution, and 10 pmol of each primer. .. The negative control consisted of RNA isolated from peripheral blood mononuclear cells from donors and the positive control was RNA isolated from the Jurkat cell line.

    Article Title: Two TonB Systems in Actinobacillus pleuropneumoniae: Their Roles in Iron Acquisition and Virulence
    Article Snippet: .. Negative control experiments to test for the presence of contaminating DNA were performed by incubating the RT-PCR mixture (prior to the addition of enzyme) with 4 μl of RNase A (100 mg/ml) for 15 min at 37°C or by using HotStart Taq (Qiagen) instead of the supplied enzyme mixture, thus eliminating the reverse transcriptase step. .. tonB1 was identified in a cosmid library of A. pleuropneumoniae serotype 1 by using a probe based on the published partial sequence , and tonB1 and associated genes were sequenced directly from the clone.

    Agarose Gel Electrophoresis:

    Article Title: Streptococcus pneumoniae induced c-Jun-N-terminal kinase- and AP-1 -dependent IL-8 release by lung epithelial BEAS-2B cells
    Article Snippet: After proteinase K digestion (1 μg/8 μl for 6 h at 37°C and 6 h at 65°) DNA was extracted using a PCR purification kit (Qiagen, Hilden, Germany). il8 promoter DNA was amplified by PCR using Hotstart Taq (Qiagen) polymerase. .. The PCR conditions were 95°C for 15 min, 33 – 35 cycles of 94°C for 20 s, 60°C for 20 s, 72°C for 20 s. PCR products were separated by agarose gel electrophoresis and detected by ethidium bromide staining.

    Activation Assay:

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: .. Followed by Hotstart Taq (Qiagen) activation at 95°C for 15 min, the first round of PCR was performed for 50 cycles of 94°C for 30s, 56°C for 30s, 72°C for 55s, and final extension at 72°C for 10 min. Primers for amplification were 5′ MsVHE (5′-GGGAATTCGAGGTGCAGCTGCAGGAGTCTGG-3′) and 3′ Cμ outer. .. 2 µl of unpurified first round PCR product was used for the seminested second round PCR, starting with Hotstart Taq (Qiagen) activation at 95°C for 15 min.

    Article Title: Carriers of the Complex Allele HFE c.[187C > G;340+4T > C] Have Increased Risk of Iron Overload in São Miguel Island Population (Azores, Portugal)
    Article Snippet: Each 20 μl PCR amplification reaction contained 100 ng of genomic DNA, 10 μM primers, 200 μM dNTPs (Promega), 25 nM MgCl2 (Qiagen), 1x Q-Solution (Qiagen), 1x buffer (Qiagen), 5 U of HotStart Taq (Qiagen), and sterile water. .. The PCR started with an enzyme activation step at 95°C for 15 min, then 40 cycles at 94°C for 30s, 56.5°C for 30s and 72°C for 1 min, followed by a final extension step at 72°C for 10 min. Amplification products were purified using the QIAquick PCR Purification kit (Qiagen), according to the manufacturer’s instructions.

    Article Title: Microbial symbionts regulate the primary Ig repertoire
    Article Snippet: .. 2 µl of unpurified first round PCR product was used for the seminested second round PCR, starting with Hotstart Taq (Qiagen) activation at 95°C for 15 min. .. The reactions were then performed for 50 cycles of 94°C for 30 s, 45°C for 30 s, 72°C for 45 s, and final extension at 72°C for 10 min. Primers for amplification were 5′ MsVHE and 3′ Cμ inner (5′-AGGGGGAAGACATTTGGGAAGGAC-3′; ).

    Article Title: Cold-sensitive mutants of Taq DNA polymerase provide a hot start for PCR
    Article Snippet: The wax melts at the first PCR heat step, causing mixing of all reaction components and activation of the DNA polymerase ( – ). .. Some enzyme preparations, such as Amplitaq Gold (Perkin Elmer and Roche Molecular Systems) and Hotstart Taq (Qiagen) are chemically inactivated and can be reactivated by heating in a special preincubation step ( ).

    Article Title: Diagnosis of Human Leptospirosis in a Clinical Setting: Real-Time PCR High Resolution Melting Analysis for Detection of Leptospira at the Onset of Disease
    Article Snippet: The first PCR reaction contained 5 μl of bacterial DNA, 10 μM primers A and B, 100 μM dNTPs (Promega), 25 nM MgCl2 (Qiagen), 1X Q-Solution (Qiagen), 1X buffer (Qiagen), 5 U of HotStart Taq (Qiagen) and RNase-free water to a final volume of 50 μl. .. The PCR programme started with an enzyme activation step at 95 °C for 15 minutes; proceeded with 30 cycles of 94 °C for 1 minute, 63 °C for 1 minute and 72 °C for 1 minute; and ended with a final extension step at 72 °C for 10 minutes.

    High Throughput Screening Assay:

    Article Title: Evidence that the Essential Response Regulator YycF in Streptococcus pneumoniae Modulates Expression of Fatty Acid Biosynthesis Genes and Alters Membrane Composition †
    Article Snippet: All PCRs were performed with HotStart Taq (QIAGEN) and a Primus-HT PCR machine (MWG Biotech) according to the following parameters: 95°C for 15 min, followed by 30 cycles of 95°C for 30 s, 50°C for 30 s, and 72°C for 120 s, with a final incubation at 72°C for 600 s. Genomic TIGR4 and R6 DNAs (2.5 ng) were used as templates, and approximately 100 pmol of each primer was used in a reaction volume of 50 μl. .. In the initial high-throughput PCR, a small percentage of reactions (approximately 5%) were either unsuccessful or gave poor yields of product.

    Staining:

    Article Title: Streptococcus pneumoniae induced c-Jun-N-terminal kinase- and AP-1 -dependent IL-8 release by lung epithelial BEAS-2B cells
    Article Snippet: After proteinase K digestion (1 μg/8 μl for 6 h at 37°C and 6 h at 65°) DNA was extracted using a PCR purification kit (Qiagen, Hilden, Germany). il8 promoter DNA was amplified by PCR using Hotstart Taq (Qiagen) polymerase. .. The PCR conditions were 95°C for 15 min, 33 – 35 cycles of 94°C for 20 s, 60°C for 20 s, 72°C for 20 s. PCR products were separated by agarose gel electrophoresis and detected by ethidium bromide staining.

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  • 99
    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
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    99
    Qiagen hotstar taq dna polymerase
    Optimisation of the PPLMD method . The y-axis represents the average background-subtracted pixel density (0–64,000). A. Introduction of SpikeTarget <t>DNA</t> 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. <t>HotStar</t> <t>Taq</t> 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.
    Hotstar Taq Dna Polymerase, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 360 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hotstar taq dna polymerase/product/Qiagen
    Average 99 stars, based on 360 article reviews
    Price from $9.99 to $1999.99
    hotstar taq dna polymerase - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

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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

    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 results showed a tendency that positive signals were improved and background signals were decreased compared to the use of HotStar Taq DNA polymerase, although statistical evidence is lacking (results not shown).

    Techniques: Negative Control