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Kapa Biosystems dntp
Dntp, supplied by Kapa Biosystems, used in various techniques. Bioz Stars score: 94/100, based on 67 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 67 article reviews
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dntp - by Bioz Stars, 2020-04
94/100 stars

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Amplification:

Article Title: ADRB2 Gene Polymorphisms and Salbutamol Responsiveness in Serbian Children with Asthma
Article Snippet: The PCR was conducted in a 50 μL reaction mixture containing: 1 × KAPA Taq Buffer A (KAPA Biosystems, Waltham, MA, USA), 0.3 mM MgCl2 , 0.2 mM each dNTP, 10 pmol of each primer, 2U of KAPA Taq DNA Polymerase (KAPA Biosystems) and approximately 300 ng of DNA. .. The obtained PCR fragments (584 bp long) were purified with PureLink PCR Purification Kit (Thermo Fisher Scientific) and sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit (Thermo Fisher Scientific) and the same primers as for the amplification.

Article Title: Characterization of nuclear microsatellite markers for Rumex bucephalophorus (Polygonaceae) using 454 sequencing 1
Article Snippet: PCR reactions, for a total volume of 25 μL, contained 2.5 μL of PCR Buffer 10×, 1 μL of MgCl2 (25 mM), 1 μL of dNTP (10 mM), 0.2 μL of KAPA Taq DNA polymerase (5 U/μL) (Kapa Biosystems, Wilmington, Massachusetts, USA), 1 μL of primer with 5′-GTTT tail (10 μM), 0.3 μL of primer with 5′-CAG or M13R tail (10 μM), and 0.5 μL of CAG or M13R with FAM, NED, PET, or VIC fluorescent label (10 μM). .. Sixteen primer pairs correctly amplified with the expected size and were run on an ABI 3730 automated sequencer (Applied Biosystems, Foster City, California, USA) using LIZ 500 as the internal lane size standard.

Article Title: ABCC2-24C > T polymorphism is associated with the response to platinum/5-Fu-based neoadjuvant chemotherapy and better clinical outcomes in advanced gastric cancer patients
Article Snippet: .. PCR amplification was performed in a total volume of 15 μL containing 50 mM KCl, 10 mM Tris-HCl (pH 8.3), 2 mM MgCl2 , 300 μM of each dNTP, 0.5 U of KAPA Taq HotStart DNA Polymerase (Kapa Biosystems), 15 pmol of each primer, and 50 ng of gDNA. .. A standard touchdown PCR protocol was used.

Article Title: Assessment of natural variation in the first pore domain of the tomato HKT1;2 transporter and characterization of mutated versions of SlHKT1;2 expressed in Xenopus laevis oocytes and via complementation of the salt sensitive athkt1;1 mutant
Article Snippet: Per reaction, 3.21 μl of MilliQ, 0.05 μl of FW primer (10 μmol/μl) and 0.25 μl RV primer (10 μmol/μl), 0.04 μl of PAL (5 U/μl) (KAPA Biosystems, Boston, USA), 0.25 μl LC Green (Idaho Technology, Salt Lake City, USA), 0.20 μl dNTP (5 mM) and 1 μl PAL buffer (KAPA Biosystems, Boston, USA) supplemented with 12.5 mM MgCl2 , were added. .. The amplification reaction started with a denaturation step of 95°C for 10 min and continued with 14 cycles of 95°C for 15 sec and 60°C for 4 min.

Article Title: A DNA 'Barcode Blitz': Rapid Digitization and Sequencing of a Natural History Collection
Article Snippet: For example, those lacking a 407 bp amplicon were assembled into a new plate and tested for the recovery of a 295 bp amplicon which enabled the generation of a 550 bp sequence record when combined with the 307 bp sequence. .. Each PCR reaction included 2 µL of DNA, 6.25 µL of 10% D-(+)-trehalose dihydrate (Fluka Analytical), 2 µL of Hyclone ultra-pure water (Thermo Scientific), 1.25 µL of 10× PlatinumTaq buffer (Invitrogen), 0.625 µL of 50 mM MgCl2 (Invitrogen), 0.125 µL of each primer, 0.0625 µL of 10 mM dNTP (KAPA Biosystems), and 0.060 µL of 5 U/µL PlatinumTaq DNA Polymerase (Invitrogen) for a total reaction volume of 12.5 µL.

Article Title: Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments
Article Snippet: Paragraph title: PCR amplification of the 18S rRNA gene ... Each 10 μl PCR mixture contained 1x Go Taq PCR buffer (KAPA Biosystems), 3.75 mM MgCl2 , 400 μM of each dNTP, 0.5 μM 18SiF primer, 0.5 μM 18SiR primer, 0.2 μM probe and 1U/reaction Kapa DNA polymerase (KAPA Biosystems).

Article Title: Molecular discrimination of Opisthorchis-like eggs from residents in a rural community of central Thailand
Article Snippet: The PCR assay was performed in a total volume of 50 μl, consisting of DNA template, 25 pmol of each primer, 200 μM dNTP, 2 mM of MgCl2 , 1X buffer, 1X enhancer, and 1 unit of KAPA2G Robust Hotstart polymerase (5U/μl) (KAPA Biosystems). .. The PCR products were amplified in the Mastercycle personal (Bio-Rad).

Article Title: Differential Preference of Burkholderia and Mesorhizobium to pH and Soil Types in the Core Cape Subregion, South Africa
Article Snippet: Paragraph title: 2.2. DNA Extraction, Amplification and Sequencing ... Each PCR reaction had a total volume of 25 µL: comprising 19.92 µL of water, 2 µL of 10× buffer (Buffer A) that contained 1.5 mM Mg2+ , 0.4 µL of 10 mM dNTP, 0.8 µL each of forward and reverse primers (10 µM), 0.08 µL of Taq polymerase (Kapa Biosystems, Cape Town, South Africa) and 1 µL of template DNA.

Article Title: Erosive processes after tectonic uplift stimulate vicariant and adaptive speciation: evolution in an Afrotemperate-endemic paper daisy genus
Article Snippet: For the chloroplast genome, the trnT-trnL spacer was amplified using the primers ‘trna’ and ‘trnb’ of [ ] and the psbA-trnH spacer was amplified with the trnH-R and psbA-F primers of [ ]. .. Reaction mixtures consisted of 12.8 μl nuclease-free H2 O, 2.5 μl of 10x buffer (Kapa Biosystems Inc., MA, USA), 1.5 μl of 25 μM MgCl2 , 1 μl dNTP mix at 0.2 μM each dNTP, 0.5 μl DMSO, 1.25 μl of each primer at 10 μM, 0.2 μl of Taq DNA polymerase (Kapa Biosystems Inc., MA, USA) and 4 μl of template DNA at various dilutions.

Article Title: Polymorphisms of the Homologous Recombination Gene RAD51 in Keratoconus and Fuchs Endothelial Corneal Dystrophy
Article Snippet: The total reaction volume of 20 μ L contained 50 ng of genomic DNA, 1 × Kapa Taq Ready Mix containing KapaTag DNA polymerase (0.025 U/μ L), reaction buffer with MgCl2 and 0.2 mM each dNTP (Kapa Biosystems, Woburn, MA, USA), and 0.25 μ M of each primer (Metabion, Martinsried, Germany). .. 131-bp length fragments containing two SNPs were amplified using the following primers: forward 5′-TGG GAA CTG CAA CTC ATC TGG-3′ and reverse 5′-GCT CCG ACT CCC CGC CGG-3′.

Article Title: HLA-A*0206 with TLR3 Polymorphisms Exerts More than Additive Effects in Stevens-Johnson Syndrome with Severe Ocular Surface Complications
Article Snippet: For direct sequencing, PCR amplification was conducted with AmpliTaq Gold DNA Polymerase (Applied Biosystems) for 35 cycles at 94°C for 1 min, annealing at 60°C for 1 min, and 72°C for 1 min on a commercial PCR machine (GeneAmp; Perkin-Elmer Applied Biosystems). .. Multiplex PCR was performed in 10 µl of Multiplex PCR buffer containing 25 ng genomic DNA, 25 nM of each multiplex primer mix, 200 µM of each dNTP, 2.25 mM MgCl2 , and 0.4 U KAPA2G Fast HotStart DNA polymerase (Kapa Biosystems).

Article Title: Microbiome composition and geochemical characteristics of deep subsurface high-pressure environment, Pyhäsalmi mine Finland
Article Snippet: .. The first step of PCR amplification was performed in 10 μL and second step in 50 μL reactions containing 1 × KAPA Fidelity buffer (Kapa Biosystems), 0.3 mM final concentration of dNTP, 6 pmol of each primer in 10μL reaction and 25 pmol in 50 μL reaction, 1 unit of KAPA Hifi polymeraze enzyme (Kapa Biosystems) and 1μL in 10 μL reaction, and 2 μL in 50 μL reaction of template. .. The PCR program for both PCR steps consisted of an initial denaturation step at 98°C for 5min, 39 cycles of 20 s at 98°C, 15 s at 53°C (for archaea and fungi at 50°C), and 30 s at 72°C.

Electrophoresis:

Article Title: ADRB2 Gene Polymorphisms and Salbutamol Responsiveness in Serbian Children with Asthma
Article Snippet: The PCR was conducted in a 50 μL reaction mixture containing: 1 × KAPA Taq Buffer A (KAPA Biosystems, Waltham, MA, USA), 0.3 mM MgCl2 , 0.2 mM each dNTP, 10 pmol of each primer, 2U of KAPA Taq DNA Polymerase (KAPA Biosystems) and approximately 300 ng of DNA. .. Products of sequencing reactions were analyzed by capillary electrophoresis on an ABI PRISM® 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA), and sequencing analysis software (Applied Biosystems).

Article Title: ABCC2-24C > T polymorphism is associated with the response to platinum/5-Fu-based neoadjuvant chemotherapy and better clinical outcomes in advanced gastric cancer patients
Article Snippet: PCR amplification was performed in a total volume of 15 μL containing 50 mM KCl, 10 mM Tris-HCl (pH 8.3), 2 mM MgCl2 , 300 μM of each dNTP, 0.5 U of KAPA Taq HotStart DNA Polymerase (Kapa Biosystems), 15 pmol of each primer, and 50 ng of gDNA. .. Capillary electrophoresis was performed on an ABI® 3130 Genetic Analyzer (Life Technologies, Inc.) and data were collected using GeneMapper® version 4.0 software (Life Technologies, Inc.).

Article Title: Differential Preference of Burkholderia and Mesorhizobium to pH and Soil Types in the Core Cape Subregion, South Africa
Article Snippet: Each PCR reaction had a total volume of 25 µL: comprising 19.92 µL of water, 2 µL of 10× buffer (Buffer A) that contained 1.5 mM Mg2+ , 0.4 µL of 10 mM dNTP, 0.8 µL each of forward and reverse primers (10 µM), 0.08 µL of Taq polymerase (Kapa Biosystems, Cape Town, South Africa) and 1 µL of template DNA. .. PCR products were loaded onto ethidium bromide agarose gels (1%) and subjected to electrophoresis using 0.5× Tris Borat EDTA (TBE).

Touchdown PCR:

Article Title: ABCC2-24C > T polymorphism is associated with the response to platinum/5-Fu-based neoadjuvant chemotherapy and better clinical outcomes in advanced gastric cancer patients
Article Snippet: PCR amplification was performed in a total volume of 15 μL containing 50 mM KCl, 10 mM Tris-HCl (pH 8.3), 2 mM MgCl2 , 300 μM of each dNTP, 0.5 U of KAPA Taq HotStart DNA Polymerase (Kapa Biosystems), 15 pmol of each primer, and 50 ng of gDNA. .. A standard touchdown PCR protocol was used.

Modification:

Article Title: A DNA 'Barcode Blitz': Rapid Digitization and Sequencing of a Natural History Collection
Article Snippet: Each PCR reaction included 2 µL of DNA, 6.25 µL of 10% D-(+)-trehalose dihydrate (Fluka Analytical), 2 µL of Hyclone ultra-pure water (Thermo Scientific), 1.25 µL of 10× PlatinumTaq buffer (Invitrogen), 0.625 µL of 50 mM MgCl2 (Invitrogen), 0.125 µL of each primer, 0.0625 µL of 10 mM dNTP (KAPA Biosystems), and 0.060 µL of 5 U/µL PlatinumTaq DNA Polymerase (Invitrogen) for a total reaction volume of 12.5 µL. .. Cycle sequencing was performed using a modified BigDye 3.1 Terminator (Applied Biosystems) protocol .

Article Title: Differential Preference of Burkholderia and Mesorhizobium to pH and Soil Types in the Core Cape Subregion, South Africa
Article Snippet: DNA Extraction, Amplification and Sequencing DNA was extracted using a modified version [ ] of the cetyl trimethylammonium bromide (CTAB) DNA Extraction protocol [ ]. .. Each PCR reaction had a total volume of 25 µL: comprising 19.92 µL of water, 2 µL of 10× buffer (Buffer A) that contained 1.5 mM Mg2+ , 0.4 µL of 10 mM dNTP, 0.8 µL each of forward and reverse primers (10 µM), 0.08 µL of Taq polymerase (Kapa Biosystems, Cape Town, South Africa) and 1 µL of template DNA.

Article Title: Erosive processes after tectonic uplift stimulate vicariant and adaptive speciation: evolution in an Afrotemperate-endemic paper daisy genus
Article Snippet: DNA extraction and sequencing Total genomic DNA was isolated from silica-dried, field-sampled material using the CTAB extraction protocol of [ ] modified according to [ ], while the Qiagen DNeasy plant extraction-kit (Qiagen Sciences, Valencia, California, U.S.A.) was used for herbarium material. .. Reaction mixtures consisted of 12.8 μl nuclease-free H2 O, 2.5 μl of 10x buffer (Kapa Biosystems Inc., MA, USA), 1.5 μl of 25 μM MgCl2 , 1 μl dNTP mix at 0.2 μM each dNTP, 0.5 μl DMSO, 1.25 μl of each primer at 10 μM, 0.2 μl of Taq DNA polymerase (Kapa Biosystems Inc., MA, USA) and 4 μl of template DNA at various dilutions.

Article Title: Microbiome composition and geochemical characteristics of deep subsurface high-pressure environment, Pyhäsalmi mine Finland
Article Snippet: Then, a second PCR with tagged ARC344f (Bano et al., ) and Ar774r primers (modified from Barns et al., ) was used to produce the tagged product for sequencing (covering the V3-V4 variable areas). .. The first step of PCR amplification was performed in 10 μL and second step in 50 μL reactions containing 1 × KAPA Fidelity buffer (Kapa Biosystems), 0.3 mM final concentration of dNTP, 6 pmol of each primer in 10μL reaction and 25 pmol in 50 μL reaction, 1 unit of KAPA Hifi polymeraze enzyme (Kapa Biosystems) and 1μL in 10 μL reaction, and 2 μL in 50 μL reaction of template.

Countercurrent Chromatography:

Article Title: Polymorphisms of the Homologous Recombination Gene RAD51 in Keratoconus and Fuchs Endothelial Corneal Dystrophy
Article Snippet: The total reaction volume of 20 μ L contained 50 ng of genomic DNA, 1 × Kapa Taq Ready Mix containing KapaTag DNA polymerase (0.025 U/μ L), reaction buffer with MgCl2 and 0.2 mM each dNTP (Kapa Biosystems, Woburn, MA, USA), and 0.25 μ M of each primer (Metabion, Martinsried, Germany). .. 131-bp length fragments containing two SNPs were amplified using the following primers: forward 5′-TGG GAA CTG CAA CTC ATC TGG-3′ and reverse 5′-GCT CCG ACT CCC CGC CGG-3′.

Genotyping Assay:

Article Title: ABCC2-24C > T polymorphism is associated with the response to platinum/5-Fu-based neoadjuvant chemotherapy and better clinical outcomes in advanced gastric cancer patients
Article Snippet: SNP genotyping Single nucleotide variant genotyping was performed using a SNaPShot™ SNP multiplex genotyping assay (ABI PRISM® SNaPshot™ Multiplex Kit, Life Technologies, Grand Island, NY, USA) according to the manufacturer's instructions. .. PCR amplification was performed in a total volume of 15 μL containing 50 mM KCl, 10 mM Tris-HCl (pH 8.3), 2 mM MgCl2 , 300 μM of each dNTP, 0.5 U of KAPA Taq HotStart DNA Polymerase (Kapa Biosystems), 15 pmol of each primer, and 50 ng of gDNA.

Digital PCR:

Article Title: Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments
Article Snippet: Absolute numbers of Cryptosporidium oocysts in these standards were determined using droplet digital PCR (ddPCR) at the 18S locus using the same primer set and these ddPCR calibrated standards were used for qPCR as previously described [ ]. .. Each 10 μl PCR mixture contained 1x Go Taq PCR buffer (KAPA Biosystems), 3.75 mM MgCl2 , 400 μM of each dNTP, 0.5 μM 18SiF primer, 0.5 μM 18SiR primer, 0.2 μM probe and 1U/reaction Kapa DNA polymerase (KAPA Biosystems).

Polymerase Chain Reaction:

Article Title: ADRB2 Gene Polymorphisms and Salbutamol Responsiveness in Serbian Children with Asthma
Article Snippet: .. The PCR was conducted in a 50 μL reaction mixture containing: 1 × KAPA Taq Buffer A (KAPA Biosystems, Waltham, MA, USA), 0.3 mM MgCl2 , 0.2 mM each dNTP, 10 pmol of each primer, 2U of KAPA Taq DNA Polymerase (KAPA Biosystems) and approximately 300 ng of DNA. ..

Article Title: Characterization of nuclear microsatellite markers for Rumex bucephalophorus (Polygonaceae) using 454 sequencing 1
Article Snippet: .. PCR reactions, for a total volume of 25 μL, contained 2.5 μL of PCR Buffer 10×, 1 μL of MgCl2 (25 mM), 1 μL of dNTP (10 mM), 0.2 μL of KAPA Taq DNA polymerase (5 U/μL) (Kapa Biosystems, Wilmington, Massachusetts, USA), 1 μL of primer with 5′-GTTT tail (10 μM), 0.3 μL of primer with 5′-CAG or M13R tail (10 μM), and 0.5 μL of CAG or M13R with FAM, NED, PET, or VIC fluorescent label (10 μM). .. Sixteen primer pairs correctly amplified with the expected size and were run on an ABI 3730 automated sequencer (Applied Biosystems, Foster City, California, USA) using LIZ 500 as the internal lane size standard.

Article Title: ABCC2-24C > T polymorphism is associated with the response to platinum/5-Fu-based neoadjuvant chemotherapy and better clinical outcomes in advanced gastric cancer patients
Article Snippet: .. PCR amplification was performed in a total volume of 15 μL containing 50 mM KCl, 10 mM Tris-HCl (pH 8.3), 2 mM MgCl2 , 300 μM of each dNTP, 0.5 U of KAPA Taq HotStart DNA Polymerase (Kapa Biosystems), 15 pmol of each primer, and 50 ng of gDNA. .. A standard touchdown PCR protocol was used.

Article Title: Assessment of natural variation in the first pore domain of the tomato HKT1;2 transporter and characterization of mutated versions of SlHKT1;2 expressed in Xenopus laevis oocytes and via complementation of the salt sensitive athkt1;1 mutant
Article Snippet: Unlabeled probes were blocked at the 3′ end to prevent extension in PCR reactions and designed to anneal over an SNP of interest. .. Per reaction, 3.21 μl of MilliQ, 0.05 μl of FW primer (10 μmol/μl) and 0.25 μl RV primer (10 μmol/μl), 0.04 μl of PAL (5 U/μl) (KAPA Biosystems, Boston, USA), 0.25 μl LC Green (Idaho Technology, Salt Lake City, USA), 0.20 μl dNTP (5 mM) and 1 μl PAL buffer (KAPA Biosystems, Boston, USA) supplemented with 12.5 mM MgCl2 , were added.

Article Title: A DNA 'Barcode Blitz': Rapid Digitization and Sequencing of a Natural History Collection
Article Snippet: .. Each PCR reaction included 2 µL of DNA, 6.25 µL of 10% D-(+)-trehalose dihydrate (Fluka Analytical), 2 µL of Hyclone ultra-pure water (Thermo Scientific), 1.25 µL of 10× PlatinumTaq buffer (Invitrogen), 0.625 µL of 50 mM MgCl2 (Invitrogen), 0.125 µL of each primer, 0.0625 µL of 10 mM dNTP (KAPA Biosystems), and 0.060 µL of 5 U/µL PlatinumTaq DNA Polymerase (Invitrogen) for a total reaction volume of 12.5 µL. ..

Article Title: Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments
Article Snippet: .. Each 10 μl PCR mixture contained 1x Go Taq PCR buffer (KAPA Biosystems), 3.75 mM MgCl2 , 400 μM of each dNTP, 0.5 μM 18SiF primer, 0.5 μM 18SiR primer, 0.2 μM probe and 1U/reaction Kapa DNA polymerase (KAPA Biosystems). .. The PCR cycling conditions consisted of one pre-melt cycle at 95°C for 6 min and then 50 cycles of 94°C for 20 sec and 60°C for 90 sec.

Article Title: Molecular discrimination of Opisthorchis-like eggs from residents in a rural community of central Thailand
Article Snippet: .. The PCR assay was performed in a total volume of 50 μl, consisting of DNA template, 25 pmol of each primer, 200 μM dNTP, 2 mM of MgCl2 , 1X buffer, 1X enhancer, and 1 unit of KAPA2G Robust Hotstart polymerase (5U/μl) (KAPA Biosystems). .. The PCR products were amplified in the Mastercycle personal (Bio-Rad).

Article Title: Differential Preference of Burkholderia and Mesorhizobium to pH and Soil Types in the Core Cape Subregion, South Africa
Article Snippet: .. Each PCR reaction had a total volume of 25 µL: comprising 19.92 µL of water, 2 µL of 10× buffer (Buffer A) that contained 1.5 mM Mg2+ , 0.4 µL of 10 mM dNTP, 0.8 µL each of forward and reverse primers (10 µM), 0.08 µL of Taq polymerase (Kapa Biosystems, Cape Town, South Africa) and 1 µL of template DNA. .. All DNA regions were amplified according to the reaction conditions described by the authors of the primers, i.e. Weisburg and co-workers [ ] for 16S rRNA, Gaunt and co-workers [ ] for recA and Haukka and co-workers [ ] for nodA .

Article Title: Erosive processes after tectonic uplift stimulate vicariant and adaptive speciation: evolution in an Afrotemperate-endemic paper daisy genus
Article Snippet: PCR was performed in an Applied Biosystems 2720 thermal cycler (Applied Biosystems CA, USA) with the following thermal profile: initial denaturation of two minutes at 94°C; 35 cycles consisting of 94°C for 45 sec, 52°C for 45 sec (annealing) and 72°C for two min (extension); and a final extension step of 72°C for eight min. .. Reaction mixtures consisted of 12.8 μl nuclease-free H2 O, 2.5 μl of 10x buffer (Kapa Biosystems Inc., MA, USA), 1.5 μl of 25 μM MgCl2 , 1 μl dNTP mix at 0.2 μM each dNTP, 0.5 μl DMSO, 1.25 μl of each primer at 10 μM, 0.2 μl of Taq DNA polymerase (Kapa Biosystems Inc., MA, USA) and 4 μl of template DNA at various dilutions.

Article Title: Polymorphisms of the Homologous Recombination Gene RAD51 in Keratoconus and Fuchs Endothelial Corneal Dystrophy
Article Snippet: SNP Genotyping The c.-61G > T and c.-98G > C polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. .. The total reaction volume of 20 μ L contained 50 ng of genomic DNA, 1 × Kapa Taq Ready Mix containing KapaTag DNA polymerase (0.025 U/μ L), reaction buffer with MgCl2 and 0.2 mM each dNTP (Kapa Biosystems, Woburn, MA, USA), and 0.25 μ M of each primer (Metabion, Martinsried, Germany).

Article Title: HLA-A*0206 with TLR3 Polymorphisms Exerts More than Additive Effects in Stevens-Johnson Syndrome with Severe Ocular Surface Complications
Article Snippet: .. Multiplex PCR was performed in 10 µl of Multiplex PCR buffer containing 25 ng genomic DNA, 25 nM of each multiplex primer mix, 200 µM of each dNTP, 2.25 mM MgCl2 , and 0.4 U KAPA2G Fast HotStart DNA polymerase (Kapa Biosystems). .. HLA-A Genotyping For HLA-A genotyping, we performed polymerase chain reaction amplification followed by hybridization with sequence-specific oligonucleotide probes (PCR-SSO) using commercial bead-based typing kits (WAK Flow, Wakunaga, Hiroshima, Japan), as described previously , .

Article Title: Microbiome composition and geochemical characteristics of deep subsurface high-pressure environment, Pyhäsalmi mine Finland
Article Snippet: .. The first step of PCR amplification was performed in 10 μL and second step in 50 μL reactions containing 1 × KAPA Fidelity buffer (Kapa Biosystems), 0.3 mM final concentration of dNTP, 6 pmol of each primer in 10μL reaction and 25 pmol in 50 μL reaction, 1 unit of KAPA Hifi polymeraze enzyme (Kapa Biosystems) and 1μL in 10 μL reaction, and 2 μL in 50 μL reaction of template. .. The PCR program for both PCR steps consisted of an initial denaturation step at 98°C for 5min, 39 cycles of 20 s at 98°C, 15 s at 53°C (for archaea and fungi at 50°C), and 30 s at 72°C.

Cellular Antioxidant Activity Assay:

Article Title: Polymorphisms of the Homologous Recombination Gene RAD51 in Keratoconus and Fuchs Endothelial Corneal Dystrophy
Article Snippet: The total reaction volume of 20 μ L contained 50 ng of genomic DNA, 1 × Kapa Taq Ready Mix containing KapaTag DNA polymerase (0.025 U/μ L), reaction buffer with MgCl2 and 0.2 mM each dNTP (Kapa Biosystems, Woburn, MA, USA), and 0.25 μ M of each primer (Metabion, Martinsried, Germany). .. 131-bp length fragments containing two SNPs were amplified using the following primers: forward 5′-TGG GAA CTG CAA CTC ATC TGG-3′ and reverse 5′-GCT CCG ACT CCC CGC CGG-3′.

DNA Extraction:

Article Title: Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments
Article Snippet: DNA was extracted from this stock using a Powersoil DNA extraction kit (MO BIO, Carlsbad, California, USA). .. Each 10 μl PCR mixture contained 1x Go Taq PCR buffer (KAPA Biosystems), 3.75 mM MgCl2 , 400 μM of each dNTP, 0.5 μM 18SiF primer, 0.5 μM 18SiR primer, 0.2 μM probe and 1U/reaction Kapa DNA polymerase (KAPA Biosystems).

Article Title: Differential Preference of Burkholderia and Mesorhizobium to pH and Soil Types in the Core Cape Subregion, South Africa
Article Snippet: Paragraph title: 2.2. DNA Extraction, Amplification and Sequencing ... Each PCR reaction had a total volume of 25 µL: comprising 19.92 µL of water, 2 µL of 10× buffer (Buffer A) that contained 1.5 mM Mg2+ , 0.4 µL of 10 mM dNTP, 0.8 µL each of forward and reverse primers (10 µM), 0.08 µL of Taq polymerase (Kapa Biosystems, Cape Town, South Africa) and 1 µL of template DNA.

Article Title: Erosive processes after tectonic uplift stimulate vicariant and adaptive speciation: evolution in an Afrotemperate-endemic paper daisy genus
Article Snippet: Paragraph title: DNA extraction and sequencing ... Reaction mixtures consisted of 12.8 μl nuclease-free H2 O, 2.5 μl of 10x buffer (Kapa Biosystems Inc., MA, USA), 1.5 μl of 25 μM MgCl2 , 1 μl dNTP mix at 0.2 μM each dNTP, 0.5 μl DMSO, 1.25 μl of each primer at 10 μM, 0.2 μl of Taq DNA polymerase (Kapa Biosystems Inc., MA, USA) and 4 μl of template DNA at various dilutions.

Isolation:

Article Title: Erosive processes after tectonic uplift stimulate vicariant and adaptive speciation: evolution in an Afrotemperate-endemic paper daisy genus
Article Snippet: DNA extraction and sequencing Total genomic DNA was isolated from silica-dried, field-sampled material using the CTAB extraction protocol of [ ] modified according to [ ], while the Qiagen DNeasy plant extraction-kit (Qiagen Sciences, Valencia, California, U.S.A.) was used for herbarium material. .. Reaction mixtures consisted of 12.8 μl nuclease-free H2 O, 2.5 μl of 10x buffer (Kapa Biosystems Inc., MA, USA), 1.5 μl of 25 μM MgCl2 , 1 μl dNTP mix at 0.2 μM each dNTP, 0.5 μl DMSO, 1.25 μl of each primer at 10 μM, 0.2 μl of Taq DNA polymerase (Kapa Biosystems Inc., MA, USA) and 4 μl of template DNA at various dilutions.

Article Title: HLA-A*0206 with TLR3 Polymorphisms Exerts More than Additive Effects in Stevens-Johnson Syndrome with Severe Ocular Surface Complications
Article Snippet: TLR3 SNPs Genotyping Genomic DNA was isolated from human peripheral blood at SRL Inc. (Tokyo, Japan). .. Multiplex PCR was performed in 10 µl of Multiplex PCR buffer containing 25 ng genomic DNA, 25 nM of each multiplex primer mix, 200 µM of each dNTP, 2.25 mM MgCl2 , and 0.4 U KAPA2G Fast HotStart DNA polymerase (Kapa Biosystems).

AST Assay:

Article Title: Erosive processes after tectonic uplift stimulate vicariant and adaptive speciation: evolution in an Afrotemperate-endemic paper daisy genus
Article Snippet: The 3′ end of the external transcribed spacer (ETS) of nuclear ribosomal DNA was amplified using the primers 18S-ETS [ ] and AST-1 [ ] while the associated ITS1 and ITS2 introns and the intervening 5.8S ribosomal gene were amplified as a unit using the ITS4 and ITS5 primers of [ ]. .. Reaction mixtures consisted of 12.8 μl nuclease-free H2 O, 2.5 μl of 10x buffer (Kapa Biosystems Inc., MA, USA), 1.5 μl of 25 μM MgCl2 , 1 μl dNTP mix at 0.2 μM each dNTP, 0.5 μl DMSO, 1.25 μl of each primer at 10 μM, 0.2 μl of Taq DNA polymerase (Kapa Biosystems Inc., MA, USA) and 4 μl of template DNA at various dilutions.

Size-exclusion Chromatography:

Article Title: Assessment of natural variation in the first pore domain of the tomato HKT1;2 transporter and characterization of mutated versions of SlHKT1;2 expressed in Xenopus laevis oocytes and via complementation of the salt sensitive athkt1;1 mutant
Article Snippet: Per reaction, 3.21 μl of MilliQ, 0.05 μl of FW primer (10 μmol/μl) and 0.25 μl RV primer (10 μmol/μl), 0.04 μl of PAL (5 U/μl) (KAPA Biosystems, Boston, USA), 0.25 μl LC Green (Idaho Technology, Salt Lake City, USA), 0.20 μl dNTP (5 mM) and 1 μl PAL buffer (KAPA Biosystems, Boston, USA) supplemented with 12.5 mM MgCl2 , were added. .. The amplification reaction started with a denaturation step of 95°C for 10 min and continued with 14 cycles of 95°C for 15 sec and 60°C for 4 min.

Article Title: Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments
Article Snippet: Each 10 μl PCR mixture contained 1x Go Taq PCR buffer (KAPA Biosystems), 3.75 mM MgCl2 , 400 μM of each dNTP, 0.5 μM 18SiF primer, 0.5 μM 18SiR primer, 0.2 μM probe and 1U/reaction Kapa DNA polymerase (KAPA Biosystems). .. The PCR cycling conditions consisted of one pre-melt cycle at 95°C for 6 min and then 50 cycles of 94°C for 20 sec and 60°C for 90 sec.

Article Title: Molecular discrimination of Opisthorchis-like eggs from residents in a rural community of central Thailand
Article Snippet: The PCR assay was performed in a total volume of 50 μl, consisting of DNA template, 25 pmol of each primer, 200 μM dNTP, 2 mM of MgCl2 , 1X buffer, 1X enhancer, and 1 unit of KAPA2G Robust Hotstart polymerase (5U/μl) (KAPA Biosystems). .. The PCR assay of COXI-Ov primer was initiated at predenaturation at 95°C for 5 min, followed by 30 cycles of denaturation at 95°C for 30 sec, annealing at 58°C for 30 sec, extension at 72°C for 30 sec, final extension was 72°C for 7 min and the holding temperature at 12°C to complete amplification.

Article Title: Erosive processes after tectonic uplift stimulate vicariant and adaptive speciation: evolution in an Afrotemperate-endemic paper daisy genus
Article Snippet: PCR was performed in an Applied Biosystems 2720 thermal cycler (Applied Biosystems CA, USA) with the following thermal profile: initial denaturation of two minutes at 94°C; 35 cycles consisting of 94°C for 45 sec, 52°C for 45 sec (annealing) and 72°C for two min (extension); and a final extension step of 72°C for eight min. .. Reaction mixtures consisted of 12.8 μl nuclease-free H2 O, 2.5 μl of 10x buffer (Kapa Biosystems Inc., MA, USA), 1.5 μl of 25 μM MgCl2 , 1 μl dNTP mix at 0.2 μM each dNTP, 0.5 μl DMSO, 1.25 μl of each primer at 10 μM, 0.2 μl of Taq DNA polymerase (Kapa Biosystems Inc., MA, USA) and 4 μl of template DNA at various dilutions.

Purification:

Article Title: ADRB2 Gene Polymorphisms and Salbutamol Responsiveness in Serbian Children with Asthma
Article Snippet: The PCR was conducted in a 50 μL reaction mixture containing: 1 × KAPA Taq Buffer A (KAPA Biosystems, Waltham, MA, USA), 0.3 mM MgCl2 , 0.2 mM each dNTP, 10 pmol of each primer, 2U of KAPA Taq DNA Polymerase (KAPA Biosystems) and approximately 300 ng of DNA. .. The obtained PCR fragments (584 bp long) were purified with PureLink PCR Purification Kit (Thermo Fisher Scientific) and sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit (Thermo Fisher Scientific) and the same primers as for the amplification.

Article Title: Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments
Article Snippet: PCR amplification of the 18S rRNA gene All samples were screened for the presence of Cryptosporidium at the 18S rRNA locus using a quantitative PCR (qPCR) previously described [ , ]. qPCR standards were Cryptosporidium oocysts (purified and haemocytometer counted), diluted to a concentration of 10,000 oocysts/μl. .. Each 10 μl PCR mixture contained 1x Go Taq PCR buffer (KAPA Biosystems), 3.75 mM MgCl2 , 400 μM of each dNTP, 0.5 μM 18SiF primer, 0.5 μM 18SiR primer, 0.2 μM probe and 1U/reaction Kapa DNA polymerase (KAPA Biosystems).

Sequencing:

Article Title: ADRB2 Gene Polymorphisms and Salbutamol Responsiveness in Serbian Children with Asthma
Article Snippet: The presence of +46A > G and +79C > G polymorphisms/variants was determined by direct sequencing of polymerase chain reaction (PCR) products obtained with the following primers: 5 ’-CTG AAT GAG GCT TCC AGG CGT-3’ and 5’-ACA ATC CAC ACC ATC AGA AT-3’. .. The PCR was conducted in a 50 μL reaction mixture containing: 1 × KAPA Taq Buffer A (KAPA Biosystems, Waltham, MA, USA), 0.3 mM MgCl2 , 0.2 mM each dNTP, 10 pmol of each primer, 2U of KAPA Taq DNA Polymerase (KAPA Biosystems) and approximately 300 ng of DNA.

Article Title: Characterization of nuclear microsatellite markers for Rumex bucephalophorus (Polygonaceae) using 454 sequencing 1
Article Snippet: The 454 sequencing reads were assembled into contigs using CAP3 at 98% sequence identity and a minimum overlap of 75 bp ( ). .. PCR reactions, for a total volume of 25 μL, contained 2.5 μL of PCR Buffer 10×, 1 μL of MgCl2 (25 mM), 1 μL of dNTP (10 mM), 0.2 μL of KAPA Taq DNA polymerase (5 U/μL) (Kapa Biosystems, Wilmington, Massachusetts, USA), 1 μL of primer with 5′-GTTT tail (10 μM), 0.3 μL of primer with 5′-CAG or M13R tail (10 μM), and 0.5 μL of CAG or M13R with FAM, NED, PET, or VIC fluorescent label (10 μM).

Article Title: A DNA 'Barcode Blitz': Rapid Digitization and Sequencing of a Natural History Collection
Article Snippet: For example, those lacking a 407 bp amplicon were assembled into a new plate and tested for the recovery of a 295 bp amplicon which enabled the generation of a 550 bp sequence record when combined with the 307 bp sequence. .. Each PCR reaction included 2 µL of DNA, 6.25 µL of 10% D-(+)-trehalose dihydrate (Fluka Analytical), 2 µL of Hyclone ultra-pure water (Thermo Scientific), 1.25 µL of 10× PlatinumTaq buffer (Invitrogen), 0.625 µL of 50 mM MgCl2 (Invitrogen), 0.125 µL of each primer, 0.0625 µL of 10 mM dNTP (KAPA Biosystems), and 0.060 µL of 5 U/µL PlatinumTaq DNA Polymerase (Invitrogen) for a total reaction volume of 12.5 µL.

Article Title: Differential Preference of Burkholderia and Mesorhizobium to pH and Soil Types in the Core Cape Subregion, South Africa
Article Snippet: Paragraph title: 2.2. DNA Extraction, Amplification and Sequencing ... Each PCR reaction had a total volume of 25 µL: comprising 19.92 µL of water, 2 µL of 10× buffer (Buffer A) that contained 1.5 mM Mg2+ , 0.4 µL of 10 mM dNTP, 0.8 µL each of forward and reverse primers (10 µM), 0.08 µL of Taq polymerase (Kapa Biosystems, Cape Town, South Africa) and 1 µL of template DNA.

Article Title: Erosive processes after tectonic uplift stimulate vicariant and adaptive speciation: evolution in an Afrotemperate-endemic paper daisy genus
Article Snippet: Paragraph title: DNA extraction and sequencing ... Reaction mixtures consisted of 12.8 μl nuclease-free H2 O, 2.5 μl of 10x buffer (Kapa Biosystems Inc., MA, USA), 1.5 μl of 25 μM MgCl2 , 1 μl dNTP mix at 0.2 μM each dNTP, 0.5 μl DMSO, 1.25 μl of each primer at 10 μM, 0.2 μl of Taq DNA polymerase (Kapa Biosystems Inc., MA, USA) and 4 μl of template DNA at various dilutions.

Article Title: HLA-A*0206 with TLR3 Polymorphisms Exerts More than Additive Effects in Stevens-Johnson Syndrome with Severe Ocular Surface Complications
Article Snippet: The PCR products were reacted with BigDye Terminator v3.1 (Applied Biosystems) and sequence reactions were resolved on an ABI PRISM 3100 Genetic Analyzer (Applied Biosystems). .. Multiplex PCR was performed in 10 µl of Multiplex PCR buffer containing 25 ng genomic DNA, 25 nM of each multiplex primer mix, 200 µM of each dNTP, 2.25 mM MgCl2 , and 0.4 U KAPA2G Fast HotStart DNA polymerase (Kapa Biosystems).

Article Title: Microbiome composition and geochemical characteristics of deep subsurface high-pressure environment, Pyhäsalmi mine Finland
Article Snippet: Then, a second PCR with tagged ARC344f (Bano et al., ) and Ar774r primers (modified from Barns et al., ) was used to produce the tagged product for sequencing (covering the V3-V4 variable areas). .. The first step of PCR amplification was performed in 10 μL and second step in 50 μL reactions containing 1 × KAPA Fidelity buffer (Kapa Biosystems), 0.3 mM final concentration of dNTP, 6 pmol of each primer in 10μL reaction and 25 pmol in 50 μL reaction, 1 unit of KAPA Hifi polymeraze enzyme (Kapa Biosystems) and 1μL in 10 μL reaction, and 2 μL in 50 μL reaction of template.

Activated Clotting Time Assay:

Article Title: Polymorphisms of the Homologous Recombination Gene RAD51 in Keratoconus and Fuchs Endothelial Corneal Dystrophy
Article Snippet: The total reaction volume of 20 μ L contained 50 ng of genomic DNA, 1 × Kapa Taq Ready Mix containing KapaTag DNA polymerase (0.025 U/μ L), reaction buffer with MgCl2 and 0.2 mM each dNTP (Kapa Biosystems, Woburn, MA, USA), and 0.25 μ M of each primer (Metabion, Martinsried, Germany). .. 131-bp length fragments containing two SNPs were amplified using the following primers: forward 5′-TGG GAA CTG CAA CTC ATC TGG-3′ and reverse 5′-GCT CCG ACT CCC CGC CGG-3′.

Software:

Article Title: ADRB2 Gene Polymorphisms and Salbutamol Responsiveness in Serbian Children with Asthma
Article Snippet: The PCR was conducted in a 50 μL reaction mixture containing: 1 × KAPA Taq Buffer A (KAPA Biosystems, Waltham, MA, USA), 0.3 mM MgCl2 , 0.2 mM each dNTP, 10 pmol of each primer, 2U of KAPA Taq DNA Polymerase (KAPA Biosystems) and approximately 300 ng of DNA. .. Products of sequencing reactions were analyzed by capillary electrophoresis on an ABI PRISM® 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA), and sequencing analysis software (Applied Biosystems).

Article Title: Characterization of nuclear microsatellite markers for Rumex bucephalophorus (Polygonaceae) using 454 sequencing 1
Article Snippet: PCR reactions, for a total volume of 25 μL, contained 2.5 μL of PCR Buffer 10×, 1 μL of MgCl2 (25 mM), 1 μL of dNTP (10 mM), 0.2 μL of KAPA Taq DNA polymerase (5 U/μL) (Kapa Biosystems, Wilmington, Massachusetts, USA), 1 μL of primer with 5′-GTTT tail (10 μM), 0.3 μL of primer with 5′-CAG or M13R tail (10 μM), and 0.5 μL of CAG or M13R with FAM, NED, PET, or VIC fluorescent label (10 μM). .. Fragment lengths were assigned to allelic classes with GeneMarker 1.71 software (SoftGenetics, State College, Pennsylvania, USA).

Article Title: ABCC2-24C > T polymorphism is associated with the response to platinum/5-Fu-based neoadjuvant chemotherapy and better clinical outcomes in advanced gastric cancer patients
Article Snippet: PCR amplification was performed in a total volume of 15 μL containing 50 mM KCl, 10 mM Tris-HCl (pH 8.3), 2 mM MgCl2 , 300 μM of each dNTP, 0.5 U of KAPA Taq HotStart DNA Polymerase (Kapa Biosystems), 15 pmol of each primer, and 50 ng of gDNA. .. Capillary electrophoresis was performed on an ABI® 3130 Genetic Analyzer (Life Technologies, Inc.) and data were collected using GeneMapper® version 4.0 software (Life Technologies, Inc.).

Article Title: Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments
Article Snippet: The 10,000 oocyst/μl DNA stock was then serially diluted to create oocyst DNA concentrations equivalent to 1000, 100, 10, 1 oocysts/μl DNA respectively to be used for standard curve generation using Rotor-Gene 6.0.14 software. .. Each 10 μl PCR mixture contained 1x Go Taq PCR buffer (KAPA Biosystems), 3.75 mM MgCl2 , 400 μM of each dNTP, 0.5 μM 18SiF primer, 0.5 μM 18SiR primer, 0.2 μM probe and 1U/reaction Kapa DNA polymerase (KAPA Biosystems).

Real-time Polymerase Chain Reaction:

Article Title: Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments
Article Snippet: Absolute numbers of Cryptosporidium oocysts in these standards were determined using droplet digital PCR (ddPCR) at the 18S locus using the same primer set and these ddPCR calibrated standards were used for qPCR as previously described [ ]. .. Each 10 μl PCR mixture contained 1x Go Taq PCR buffer (KAPA Biosystems), 3.75 mM MgCl2 , 400 μM of each dNTP, 0.5 μM 18SiF primer, 0.5 μM 18SiR primer, 0.2 μM probe and 1U/reaction Kapa DNA polymerase (KAPA Biosystems).

Multiplex Assay:

Article Title: ABCC2-24C > T polymorphism is associated with the response to platinum/5-Fu-based neoadjuvant chemotherapy and better clinical outcomes in advanced gastric cancer patients
Article Snippet: Genetic regions flanking each genetic variant were co-amplified using 2 multiplex PCR primer pools. .. PCR amplification was performed in a total volume of 15 μL containing 50 mM KCl, 10 mM Tris-HCl (pH 8.3), 2 mM MgCl2 , 300 μM of each dNTP, 0.5 U of KAPA Taq HotStart DNA Polymerase (Kapa Biosystems), 15 pmol of each primer, and 50 ng of gDNA.

Article Title: HLA-A*0206 with TLR3 Polymorphisms Exerts More than Additive Effects in Stevens-Johnson Syndrome with Severe Ocular Surface Complications
Article Snippet: .. Multiplex PCR was performed in 10 µl of Multiplex PCR buffer containing 25 ng genomic DNA, 25 nM of each multiplex primer mix, 200 µM of each dNTP, 2.25 mM MgCl2 , and 0.4 U KAPA2G Fast HotStart DNA polymerase (Kapa Biosystems). .. HLA-A Genotyping For HLA-A genotyping, we performed polymerase chain reaction amplification followed by hybridization with sequence-specific oligonucleotide probes (PCR-SSO) using commercial bead-based typing kits (WAK Flow, Wakunaga, Hiroshima, Japan), as described previously , .

Positron Emission Tomography:

Article Title: Characterization of nuclear microsatellite markers for Rumex bucephalophorus (Polygonaceae) using 454 sequencing 1
Article Snippet: .. PCR reactions, for a total volume of 25 μL, contained 2.5 μL of PCR Buffer 10×, 1 μL of MgCl2 (25 mM), 1 μL of dNTP (10 mM), 0.2 μL of KAPA Taq DNA polymerase (5 U/μL) (Kapa Biosystems, Wilmington, Massachusetts, USA), 1 μL of primer with 5′-GTTT tail (10 μM), 0.3 μL of primer with 5′-CAG or M13R tail (10 μM), and 0.5 μL of CAG or M13R with FAM, NED, PET, or VIC fluorescent label (10 μM). .. Sixteen primer pairs correctly amplified with the expected size and were run on an ABI 3730 automated sequencer (Applied Biosystems, Foster City, California, USA) using LIZ 500 as the internal lane size standard.

Produced:

Article Title: Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments
Article Snippet: Each 10 μl PCR mixture contained 1x Go Taq PCR buffer (KAPA Biosystems), 3.75 mM MgCl2 , 400 μM of each dNTP, 0.5 μM 18SiF primer, 0.5 μM 18SiR primer, 0.2 μM probe and 1U/reaction Kapa DNA polymerase (KAPA Biosystems). .. Samples that were positive by qPCR were amplified at the 18S locus using primers which produced a 611 bp product ( ) as previously described [ ] with minor modifications; the annealing temperature used in the present study was 57°C for 30 sec and the number of cycles was increased from 39 to 47 cycles for both primary and secondary reactions.

Concentration Assay:

Article Title: Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments
Article Snippet: PCR amplification of the 18S rRNA gene All samples were screened for the presence of Cryptosporidium at the 18S rRNA locus using a quantitative PCR (qPCR) previously described [ , ]. qPCR standards were Cryptosporidium oocysts (purified and haemocytometer counted), diluted to a concentration of 10,000 oocysts/μl. .. Each 10 μl PCR mixture contained 1x Go Taq PCR buffer (KAPA Biosystems), 3.75 mM MgCl2 , 400 μM of each dNTP, 0.5 μM 18SiF primer, 0.5 μM 18SiR primer, 0.2 μM probe and 1U/reaction Kapa DNA polymerase (KAPA Biosystems).

Article Title: Microbiome composition and geochemical characteristics of deep subsurface high-pressure environment, Pyhäsalmi mine Finland
Article Snippet: .. The first step of PCR amplification was performed in 10 μL and second step in 50 μL reactions containing 1 × KAPA Fidelity buffer (Kapa Biosystems), 0.3 mM final concentration of dNTP, 6 pmol of each primer in 10μL reaction and 25 pmol in 50 μL reaction, 1 unit of KAPA Hifi polymeraze enzyme (Kapa Biosystems) and 1μL in 10 μL reaction, and 2 μL in 50 μL reaction of template. .. The PCR program for both PCR steps consisted of an initial denaturation step at 98°C for 5min, 39 cycles of 20 s at 98°C, 15 s at 53°C (for archaea and fungi at 50°C), and 30 s at 72°C.

CTG Assay:

Article Title: ADRB2 Gene Polymorphisms and Salbutamol Responsiveness in Serbian Children with Asthma
Article Snippet: The presence of +46A > G and +79C > G polymorphisms/variants was determined by direct sequencing of polymerase chain reaction (PCR) products obtained with the following primers: 5 ’-CTG AAT GAG GCT TCC AGG CGT-3’ and 5’-ACA ATC CAC ACC ATC AGA AT-3’. .. The PCR was conducted in a 50 μL reaction mixture containing: 1 × KAPA Taq Buffer A (KAPA Biosystems, Waltham, MA, USA), 0.3 mM MgCl2 , 0.2 mM each dNTP, 10 pmol of each primer, 2U of KAPA Taq DNA Polymerase (KAPA Biosystems) and approximately 300 ng of DNA.

Article Title: Polymorphisms of the Homologous Recombination Gene RAD51 in Keratoconus and Fuchs Endothelial Corneal Dystrophy
Article Snippet: The total reaction volume of 20 μ L contained 50 ng of genomic DNA, 1 × Kapa Taq Ready Mix containing KapaTag DNA polymerase (0.025 U/μ L), reaction buffer with MgCl2 and 0.2 mM each dNTP (Kapa Biosystems, Woburn, MA, USA), and 0.25 μ M of each primer (Metabion, Martinsried, Germany). .. 131-bp length fragments containing two SNPs were amplified using the following primers: forward 5′-TGG GAA CTG CAA CTC ATC TGG-3′ and reverse 5′-GCT CCG ACT CCC CGC CGG-3′.

Variant Assay:

Article Title: ABCC2-24C > T polymorphism is associated with the response to platinum/5-Fu-based neoadjuvant chemotherapy and better clinical outcomes in advanced gastric cancer patients
Article Snippet: Genetic regions flanking each genetic variant were co-amplified using 2 multiplex PCR primer pools. .. PCR amplification was performed in a total volume of 15 μL containing 50 mM KCl, 10 mM Tris-HCl (pH 8.3), 2 mM MgCl2 , 300 μM of each dNTP, 0.5 U of KAPA Taq HotStart DNA Polymerase (Kapa Biosystems), 15 pmol of each primer, and 50 ng of gDNA.

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    Kapa Biosystems dntps
    Pathogenic Homozygous Recessive Variants in MCM9 Were Identified in Two Consanguineous Kurdish Families from Turkey (A and B) The MCM9 genotype is provided below each individual. WT denotes the wild-type allele. MT (for mutation) indicates either (A) MCM9 c.1732+2T > C or (B) MCM9 c. 394C > T. PCR amplification was completed with the <t>KAPA</t> HiFi HotStart PCR Kit with <t>dNTPs</t> (Kapa Biosystems) according to the manufacturer’s instructions (primers for A are 5′-TCTAGGAGGTCCCGAGATGG-3′ and 5′-CAAAGGCAGAGTGATTGCCG-3′; primers for B are 5′-GCCTGAGAGGCAAGTGAATTTAG-3′ and 5′-TACCTAAAACCAAGGATGTGGGA-3′). PCR products were sequenced at Beckman Coulter Genomics. Results were analyzed with Sequencher (Gene Codes Corporation). Sample chromatograms are shown (5′ to 3′ direction in A; 3′ to 5′ direction in B). Arrows point to the nucleotide peak of interest. Samples utilized for whole-exome sequencing (WES) are indicated by an asterisk. (C) MCM9 is encoded on chr6: 119,252,903–119,134,612 (UCSC Genome Browser hg19). The MCM9 c.1732+2T > C splice variant lies in the splice donor site of exon 9 (red arrow). The c.394C > T mutation lies near the 5′ end of exon 2 (blue arrow). Exons are indicated as full boxes (black). Alternative splice forms of MCM9 either include or exclude exon 11 (white). Introns are indicated as lines. (D) MCM9 consists of an N-terminal DNA binding domain and AAA + core domain. The c.1732+2T > C mutation is predicted to disrupt normal splicing after amino acid 577. The c.394C > T mutation causes insertion of a stop codon in the place of an arginine at position 132 (p.Arg132 ∗ ). This most likely results in a truncated protein if one is formed. The yellow box represents the arginine finger domain (RF), the pink box represents the Walker A motif (WA), and the deep green box represents the Walker B motif (WB).
    Dntps, supplied by Kapa Biosystems, used in various techniques. Bioz Stars score: 95/100, based on 92 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Pathogenic Homozygous Recessive Variants in MCM9 Were Identified in Two Consanguineous Kurdish Families from Turkey (A and B) The MCM9 genotype is provided below each individual. WT denotes the wild-type allele. MT (for mutation) indicates either (A) MCM9 c.1732+2T > C or (B) MCM9 c. 394C > T. PCR amplification was completed with the KAPA HiFi HotStart PCR Kit with dNTPs (Kapa Biosystems) according to the manufacturer’s instructions (primers for A are 5′-TCTAGGAGGTCCCGAGATGG-3′ and 5′-CAAAGGCAGAGTGATTGCCG-3′; primers for B are 5′-GCCTGAGAGGCAAGTGAATTTAG-3′ and 5′-TACCTAAAACCAAGGATGTGGGA-3′). PCR products were sequenced at Beckman Coulter Genomics. Results were analyzed with Sequencher (Gene Codes Corporation). Sample chromatograms are shown (5′ to 3′ direction in A; 3′ to 5′ direction in B). Arrows point to the nucleotide peak of interest. Samples utilized for whole-exome sequencing (WES) are indicated by an asterisk. (C) MCM9 is encoded on chr6: 119,252,903–119,134,612 (UCSC Genome Browser hg19). The MCM9 c.1732+2T > C splice variant lies in the splice donor site of exon 9 (red arrow). The c.394C > T mutation lies near the 5′ end of exon 2 (blue arrow). Exons are indicated as full boxes (black). Alternative splice forms of MCM9 either include or exclude exon 11 (white). Introns are indicated as lines. (D) MCM9 consists of an N-terminal DNA binding domain and AAA + core domain. The c.1732+2T > C mutation is predicted to disrupt normal splicing after amino acid 577. The c.394C > T mutation causes insertion of a stop codon in the place of an arginine at position 132 (p.Arg132 ∗ ). This most likely results in a truncated protein if one is formed. The yellow box represents the arginine finger domain (RF), the pink box represents the Walker A motif (WA), and the deep green box represents the Walker B motif (WB).

    Journal: American Journal of Human Genetics

    Article Title: MCM9 Mutations Are Associated with Ovarian Failure, Short Stature, and Chromosomal Instability

    doi: 10.1016/j.ajhg.2014.11.002

    Figure Lengend Snippet: Pathogenic Homozygous Recessive Variants in MCM9 Were Identified in Two Consanguineous Kurdish Families from Turkey (A and B) The MCM9 genotype is provided below each individual. WT denotes the wild-type allele. MT (for mutation) indicates either (A) MCM9 c.1732+2T > C or (B) MCM9 c. 394C > T. PCR amplification was completed with the KAPA HiFi HotStart PCR Kit with dNTPs (Kapa Biosystems) according to the manufacturer’s instructions (primers for A are 5′-TCTAGGAGGTCCCGAGATGG-3′ and 5′-CAAAGGCAGAGTGATTGCCG-3′; primers for B are 5′-GCCTGAGAGGCAAGTGAATTTAG-3′ and 5′-TACCTAAAACCAAGGATGTGGGA-3′). PCR products were sequenced at Beckman Coulter Genomics. Results were analyzed with Sequencher (Gene Codes Corporation). Sample chromatograms are shown (5′ to 3′ direction in A; 3′ to 5′ direction in B). Arrows point to the nucleotide peak of interest. Samples utilized for whole-exome sequencing (WES) are indicated by an asterisk. (C) MCM9 is encoded on chr6: 119,252,903–119,134,612 (UCSC Genome Browser hg19). The MCM9 c.1732+2T > C splice variant lies in the splice donor site of exon 9 (red arrow). The c.394C > T mutation lies near the 5′ end of exon 2 (blue arrow). Exons are indicated as full boxes (black). Alternative splice forms of MCM9 either include or exclude exon 11 (white). Introns are indicated as lines. (D) MCM9 consists of an N-terminal DNA binding domain and AAA + core domain. The c.1732+2T > C mutation is predicted to disrupt normal splicing after amino acid 577. The c.394C > T mutation causes insertion of a stop codon in the place of an arginine at position 132 (p.Arg132 ∗ ). This most likely results in a truncated protein if one is formed. The yellow box represents the arginine finger domain (RF), the pink box represents the Walker A motif (WA), and the deep green box represents the Walker B motif (WB).

    Article Snippet: Using the KAPA HiFi HotStart PCR Kit with dNTPs (Kapa Biosystems) and primers to amplify exons 7–12 (c.1151_1996), we expected to observe two previously reported wild-type products: an 846 bp product and a 700 bp product resulting from exon 11 skipping ( A; B, S3C, and A).

    Techniques: Mutagenesis, Polymerase Chain Reaction, Amplification, Sequencing, Variant Assay, Binding Assay, Western Blot