hotstart taq dna polymerase  (Qiagen)


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    Name:
    HotStarTaq Plus DNA Polymerase
    Description:
    For fast and highly specific amplification in all applications Kit contents Qiagen HotStarTaq Plus DNA Polymerase 50U 5U L 10 min at 97C 60 min at 94C Half life 2 to 4 kb min at 72C Extension Rate 5 3 Exonuclease Enzyme Activity Genomic DNA and cDNA Sample PCR Amplification Reaction Ready to load PCR Buffer Ideal for Fast and Highly Specific Amplification Includes 10x PCR Buffer 10x CoralLoad PCR Buffer 5x Q Solution 25mM MgCl2 Benefits High PCR specificity with minimal optimization Fast 5 minute enzyme activation time Ready to load PCR buffer for faster and easier handling
    Catalog Number:
    203601
    Price:
    32.8
    Category:
    HotStarTaq Plus DNA Polymerase
    Buy from Supplier


    Structured Review

    Qiagen hotstart taq dna polymerase
    HotStarTaq Plus DNA Polymerase
    For fast and highly specific amplification in all applications Kit contents Qiagen HotStarTaq Plus DNA Polymerase 50U 5U L 10 min at 97C 60 min at 94C Half life 2 to 4 kb min at 72C Extension Rate 5 3 Exonuclease Enzyme Activity Genomic DNA and cDNA Sample PCR Amplification Reaction Ready to load PCR Buffer Ideal for Fast and Highly Specific Amplification Includes 10x PCR Buffer 10x CoralLoad PCR Buffer 5x Q Solution 25mM MgCl2 Benefits High PCR specificity with minimal optimization Fast 5 minute enzyme activation time Ready to load PCR buffer for faster and easier handling
    https://www.bioz.com/result/hotstart taq dna polymerase/product/Qiagen
    Average 99 stars, based on 110 article reviews
    Price from $9.99 to $1999.99
    hotstart taq dna polymerase - by Bioz Stars, 2020-04
    99/100 stars

    Images

    1) Product Images from "Alkaline-mediated differential interaction (AMDI): A simple automatable single-nucleotide polymorphism assay"

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

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

    doi: 10.1073/pnas.041619998

    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.
    Figure Legend 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.

    Techniques Used: Polymerase Chain Reaction, Fluorescence, Amplification, Produced

    Related Articles

    Methylation Sequencing:

    Article Title: Suberanilohydroxamic acid prevents TGF-β1-induced COX-2 repression in human lung fibroblasts post-transcriptionally by TIA-1 downregulation
    Article Snippet: Paragraph title: Bisulfite sequencing ... COX -2 promoter region was amplified using 1 μM primers (primer sequences: forward 5′-GGTAGGAAATTTTATATTGGTGATT-3′ and reverse 5′-CTCACCTATATAACTAAACRCCA-3′), 2.5 mM MgCl2 , 1 μl HotStarTaq Plus DNA Polymerase (203601, Qiagen, Venlo, NL).

    Article Title: Interplay between SIN3A and STAT3 Mediates Chromatin Conformational Changes and GFAP Expression during Cellular Differentiation
    Article Snippet: Paragraph title: Bisulfite sequencing ... The PCRs were carried out with 25 ng bisulfite-treated DNA, 1 µl of HotStart Taq DNA polymerase (Qiagen), 10× supplied HotStart buffer, 4 µl of dNTP (25 mM), and 3 µl of PCR primers (5 µM).

    Clone Assay:

    Article Title: Suberanilohydroxamic acid prevents TGF-β1-induced COX-2 repression in human lung fibroblasts post-transcriptionally by TIA-1 downregulation
    Article Snippet: COX -2 promoter region was amplified using 1 μM primers (primer sequences: forward 5′-GGTAGGAAATTTTATATTGGTGATT-3′ and reverse 5′-CTCACCTATATAACTAAACRCCA-3′), 2.5 mM MgCl2 , 1 μl HotStarTaq Plus DNA Polymerase (203601, Qiagen, Venlo, NL). .. DNA was then cloned using the pGEM®-T Easy Vector System II (A1380, Promega, Madison, WI, USA).

    Article Title: Interplay between SIN3A and STAT3 Mediates Chromatin Conformational Changes and GFAP Expression during Cellular Differentiation
    Article Snippet: The PCRs were carried out with 25 ng bisulfite-treated DNA, 1 µl of HotStart Taq DNA polymerase (Qiagen), 10× supplied HotStart buffer, 4 µl of dNTP (25 mM), and 3 µl of PCR primers (5 µM). .. Several clones of each sample were verified by sequencing with the T7 universal primer.

    Article Title: Tissue-specific transcriptomics, chromosomal localization, and phylogeny of chemosensory and odorant binding proteins from the red flour beetle Tribolium castaneum reveal subgroup specificities for olfaction or more general functions
    Article Snippet: Paragraph title: Cloning of selected OBPs and CSPs open reading frames ... Hotstart Taq DNA polymerase (Qiagen, Venlo, Netherlands) was used to amplify individual transcripts.

    Article Title: Structural studies of the catalytic core of the primate foamy virus (PFV-1) integrase
    Article Snippet: Paragraph title: 2.1. Cloning and mutagenesis   ... The PFV catalytic domain (amino acids 110–309) was amplified by PCR using oligonucleotide primers 5′-CATG CATATG GGTCCTATTCTAAGACCAGATAGG-3′ and 5′-CCGCTCGAGCTAAGAGGAGG CTGGAG GGGTGGATGG-3′ and HotStart HiFi Taq DNA polymerase (Qiagen).

    Amplification:

    Article Title: Modulation of neurite outgrowth by activation of calcium-permeable kainate receptors expressed by rat nociceptive-like dorsal root ganglion neurons
    Article Snippet: Reaction mixture consisted of 1 μg of DRG or dorsal horn complementary DNA (cDNA), 10 pmol of each primer pair, 200 μM deoxynucleoside triphosphates (dNTPs), 50 mM KCl2, 10 mM Tris–HCl (pH 8.3), 1.5 mM MgCl2 , and 0.5 μl of HotStart Plus DNA polymerase (Qiagen). .. The primers used for PCR amplification, expected PCR product sizes, and their respective annealing temperatures are listed in .

    Article Title: Suberanilohydroxamic acid prevents TGF-β1-induced COX-2 repression in human lung fibroblasts post-transcriptionally by TIA-1 downregulation
    Article Snippet: .. COX -2 promoter region was amplified using 1 μM primers (primer sequences: forward 5′-GGTAGGAAATTTTATATTGGTGATT-3′ and reverse 5′-CTCACCTATATAACTAAACRCCA-3′), 2.5 mM MgCl2 , 1 μl HotStarTaq Plus DNA Polymerase (203601, Qiagen, Venlo, NL). ..

    Article Title: Effects of Upconversion Nanoparticles on Polymerase Chain Reaction
    Article Snippet: Paragraph title: Effects of UCNPs on PCR Specificity and Amplification Efficiency ... With the Qiagen HotStarTaq Plus DNA Polymerase in the Qiagen QuantiTect SYBR Green PCR Kit, the optimal concentrations of UCNPs for improving the PCR specificity were 0.5 µg/µL for the 40-nm-sized UCNPs, and 10× and 1× for the 70- and 250-nm-sized UCNPs, respectively, with a CT value of 13.44±0.07, 13.50±0.05, and 13.35±0.09, respectively (QuantiTect SYBR Green PCR Kit; 13.24±0.07 without UCNPs; ).

    Article Title: Valproic acid shows potent antitumor effect with alteration of DNA methylation in neuroblastoma
    Article Snippet: The 97 bp amplicon of THBS1 with 10 CpG dinucleotides was amplified with sense primer 5’-GGTCGGAGGAATTTTTAGGAATG-3’, and antisense primer 5’-CCTAAACTCGCAAACCAACT-3’. .. PCR Reactions were performed in triplicate in a 20 µl volume containing 1× buffer, 2U Hotstart Taq DNA polymerase (Qiagen), 250 nM of each primer, 1.5 µM SYTO-9, and 10 ng bisulfite-treated DNA template, with 2 mM final MgCl2 .

    Article Title: Structural studies of the catalytic core of the primate foamy virus (PFV-1) integrase
    Article Snippet: .. The PFV catalytic domain (amino acids 110–309) was amplified by PCR using oligonucleotide primers 5′-CATG CATATG GGTCCTATTCTAAGACCAGATAGG-3′ and 5′-CCGCTCGAGCTAAGAGGAGG CTGGAG GGGTGGATGG-3′ and HotStart HiFi Taq DNA polymerase (Qiagen). .. The PCR product was digested by Nde I and Bam HI (NEB) and cloned in pET-15b vector (Novagen) between Nde I and Xho I restriction sites.

    Article Title: Pyrosequencing versus methylation-specific PCR for assessment of MGMT methylation in tumor and blood samples of glioblastoma patients
    Article Snippet: .. MGMT methylation assessment by MSP Bisulfite-converted DNA (2 μl) was amplified using specific primers (previously described for CpG sites 74–78 ) for methylated and unmethylated DNA independently, using HotStart®Plus DNA polymerase (Qiagen, Izasa, Spain) and following the manufacturer’s instructions. .. PCR reactions (15 μl) were analyzed on a 2% agarose gel stained with ethidium bromide or Syber Safe.

    Article Title: Gene expression in the developing mouse retina by EST sequencing and microarray analysis
    Article Snippet: .. Total RNA was reverse transcribed with Superscript reverse transcriptase (Life Technologies) at 42°C. cDNA equivalent to 40 ng of total RNA was used for PCR amplification with the Hotstart Taq DNA polymerase (Qiagen) in a volume of 20 µl. ..

    Synthesized:

    Article Title: Tissue-specific transcriptomics, chromosomal localization, and phylogeny of chemosensory and odorant binding proteins from the red flour beetle Tribolium castaneum reveal subgroup specificities for olfaction or more general functions
    Article Snippet: Messenger RNA was purified with the Dynabeads purification kit (life technologies, Carlsbad, USA) and cDNA was synthesized using the Super-Script first-strand synthesis system (life technologies, Carlsbad, USA). .. Hotstart Taq DNA polymerase (Qiagen, Venlo, Netherlands) was used to amplify individual transcripts.

    TA Cloning:

    Article Title: Interplay between SIN3A and STAT3 Mediates Chromatin Conformational Changes and GFAP Expression during Cellular Differentiation
    Article Snippet: The PCRs were carried out with 25 ng bisulfite-treated DNA, 1 µl of HotStart Taq DNA polymerase (Qiagen), 10× supplied HotStart buffer, 4 µl of dNTP (25 mM), and 3 µl of PCR primers (5 µM). .. The PCRs were carried out with 25 ng bisulfite-treated DNA, 1 µl of HotStart Taq DNA polymerase (Qiagen), 10× supplied HotStart buffer, 4 µl of dNTP (25 mM), and 3 µl of PCR primers (5 µM).

    Quantitative RT-PCR:

    Article Title: Gene expression in the developing mouse retina by EST sequencing and microarray analysis
    Article Snippet: Paragraph title: Quantitative RT–PCR ... Total RNA was reverse transcribed with Superscript reverse transcriptase (Life Technologies) at 42°C. cDNA equivalent to 40 ng of total RNA was used for PCR amplification with the Hotstart Taq DNA polymerase (Qiagen) in a volume of 20 µl.

    SYBR Green Assay:

    Article Title: Effects of Upconversion Nanoparticles on Polymerase Chain Reaction
    Article Snippet: .. With the Qiagen HotStarTaq Plus DNA Polymerase in the Qiagen QuantiTect SYBR Green PCR Kit, the optimal concentrations of UCNPs for improving the PCR specificity were 0.5 µg/µL for the 40-nm-sized UCNPs, and 10× and 1× for the 70- and 250-nm-sized UCNPs, respectively, with a CT value of 13.44±0.07, 13.50±0.05, and 13.35±0.09, respectively (QuantiTect SYBR Green PCR Kit; 13.24±0.07 without UCNPs; ). .. PCR Amplification at Low Annealing Temperature At annealing temperatures ranging from 25°C to 60°C, specific PCR products were obtained, without nonspecific amplification, by addition of the 40 nm UCNP (1 µg/µL) to the PCR master mix ( ).

    Mass Spectrometry:

    Article Title: Valproic acid shows potent antitumor effect with alteration of DNA methylation in neuroblastoma
    Article Snippet: MS-HRM for RASSF1A was performed as previously described [ ]. .. PCR Reactions were performed in triplicate in a 20 µl volume containing 1× buffer, 2U Hotstart Taq DNA polymerase (Qiagen), 250 nM of each primer, 1.5 µM SYTO-9, and 10 ng bisulfite-treated DNA template, with 2 mM final MgCl2 .

    Modification:

    Article Title: Effects of Upconversion Nanoparticles on Polymerase Chain Reaction
    Article Snippet: .. The AmpliTaq Gold DNA Polymerase in the AmpliTaq Gold 360 Master Mix, and the HotStarTaq Plus DNA Polymerase in the HotStarTaq Plus Master Mix (Qiagen) were recombinant forms modified from the Thermus aquaticus (Taq ) DNA polymerase. .. Aliquots of the UCNPs solutions were added to the PCR mixtures to achieve appropriate concentrations.

    DNA Methylation Assay:

    Article Title: Suberanilohydroxamic acid prevents TGF-β1-induced COX-2 repression in human lung fibroblasts post-transcriptionally by TIA-1 downregulation
    Article Snippet: Bisulfite conversion of DNA (1 μg) was conducted with the EZ DNA Methylation™ Kit (D5001, Zymo Research, Irvine, CA, USA) following the manufacturer's instructions. .. COX -2 promoter region was amplified using 1 μM primers (primer sequences: forward 5′-GGTAGGAAATTTTATATTGGTGATT-3′ and reverse 5′-CTCACCTATATAACTAAACRCCA-3′), 2.5 mM MgCl2 , 1 μl HotStarTaq Plus DNA Polymerase (203601, Qiagen, Venlo, NL).

    Article Title: Valproic acid shows potent antitumor effect with alteration of DNA methylation in neuroblastoma
    Article Snippet: Paragraph title: DNA methylation analysis ... PCR Reactions were performed in triplicate in a 20 µl volume containing 1× buffer, 2U Hotstart Taq DNA polymerase (Qiagen), 250 nM of each primer, 1.5 µM SYTO-9, and 10 ng bisulfite-treated DNA template, with 2 mM final MgCl2 .

    Inhibition:

    Article Title: Effects of Upconversion Nanoparticles on Polymerase Chain Reaction
    Article Snippet: Complete PCR inhibition was observed when 25× to 30× UCNPs of 70 and 250 nm were added to only the AmpliTaq Gold 360 Master Mix with AmpliTaq Gold DNA Polymerase. .. With the Qiagen HotStarTaq Plus DNA Polymerase in the Qiagen QuantiTect SYBR Green PCR Kit, the optimal concentrations of UCNPs for improving the PCR specificity were 0.5 µg/µL for the 40-nm-sized UCNPs, and 10× and 1× for the 70- and 250-nm-sized UCNPs, respectively, with a CT value of 13.44±0.07, 13.50±0.05, and 13.35±0.09, respectively (QuantiTect SYBR Green PCR Kit; 13.24±0.07 without UCNPs; ).

    Sequencing:

    Article Title: Interplay between SIN3A and STAT3 Mediates Chromatin Conformational Changes and GFAP Expression during Cellular Differentiation
    Article Snippet: The PCRs were carried out with 25 ng bisulfite-treated DNA, 1 µl of HotStart Taq DNA polymerase (Qiagen), 10× supplied HotStart buffer, 4 µl of dNTP (25 mM), and 3 µl of PCR primers (5 µM). .. Several clones of each sample were verified by sequencing with the T7 universal primer.

    Article Title: Tissue-specific transcriptomics, chromosomal localization, and phylogeny of chemosensory and odorant binding proteins from the red flour beetle Tribolium castaneum reveal subgroup specificities for olfaction or more general functions
    Article Snippet: Hotstart Taq DNA polymerase (Qiagen, Venlo, Netherlands) was used to amplify individual transcripts. .. Finally the products were cloned into PCR2.1 vector (life technologies, Carlsbad, USA) and verified by sequencing.

    Article Title: Bacterial Endophytes Isolated from Plants in Natural Oil Seep Soils with Chronic Hydrocarbon Contamination
    Article Snippet: The PCR reaction was as follows: 25 μL reactions with a final concentration of 0.5 mM of the forward primer and reverse primer, 1.5 mM MgCl2 , 200 mM of each dNTP, 2.5 units of HotStarTaq Plus DNA polymerase (Qiagen, Canada). .. The resultant amplicons were purified (GenElute PCR clean-up kit, Sigma Aldrich) before being sent for Sanger sequencing at The Centre for Applied Genomics (TCAG) sequencing facility (Toronto, Canada) using 27F (5′-AGAGTTTGATYMTGGCTCAG-3′) primer.

    Recombinant:

    Article Title: Effects of Upconversion Nanoparticles on Polymerase Chain Reaction
    Article Snippet: .. The AmpliTaq Gold DNA Polymerase in the AmpliTaq Gold 360 Master Mix, and the HotStarTaq Plus DNA Polymerase in the HotStarTaq Plus Master Mix (Qiagen) were recombinant forms modified from the Thermus aquaticus (Taq ) DNA polymerase. .. Aliquots of the UCNPs solutions were added to the PCR mixtures to achieve appropriate concentrations.

    Methylation:

    Article Title: Interplay between SIN3A and STAT3 Mediates Chromatin Conformational Changes and GFAP Expression during Cellular Differentiation
    Article Snippet: Bisulfite sequencing Genomic DNA (500 ng) was bisulfite converted with the EZ DNA methylation-Gold kit (D5005; Zymo Research, Orange, CA, USA) according to the manufacturer's recommendations. .. The PCRs were carried out with 25 ng bisulfite-treated DNA, 1 µl of HotStart Taq DNA polymerase (Qiagen), 10× supplied HotStart buffer, 4 µl of dNTP (25 mM), and 3 µl of PCR primers (5 µM).

    Article Title: Valproic acid shows potent antitumor effect with alteration of DNA methylation in neuroblastoma
    Article Snippet: Methylation Sensitive High Resolution Melting (MS-HRM) technology [ ] was used to determine the methylation degree along the promoter regions of THBS-1 and RASSF1A . .. PCR Reactions were performed in triplicate in a 20 µl volume containing 1× buffer, 2U Hotstart Taq DNA polymerase (Qiagen), 250 nM of each primer, 1.5 µM SYTO-9, and 10 ng bisulfite-treated DNA template, with 2 mM final MgCl2 .

    Article Title: Pyrosequencing versus methylation-specific PCR for assessment of MGMT methylation in tumor and blood samples of glioblastoma patients
    Article Snippet: .. MGMT methylation assessment by MSP Bisulfite-converted DNA (2 μl) was amplified using specific primers (previously described for CpG sites 74–78 ) for methylated and unmethylated DNA independently, using HotStart®Plus DNA polymerase (Qiagen, Izasa, Spain) and following the manufacturer’s instructions. .. PCR reactions (15 μl) were analyzed on a 2% agarose gel stained with ethidium bromide or Syber Safe.

    Mutagenesis:

    Article Title: Structural studies of the catalytic core of the primate foamy virus (PFV-1) integrase
    Article Snippet: Paragraph title: 2.1. Cloning and mutagenesis   ... The PFV catalytic domain (amino acids 110–309) was amplified by PCR using oligonucleotide primers 5′-CATG CATATG GGTCCTATTCTAAGACCAGATAGG-3′ and 5′-CCGCTCGAGCTAAGAGGAGG CTGGAG GGGTGGATGG-3′ and HotStart HiFi Taq DNA polymerase (Qiagen).

    Isolation:

    Article Title: Bacterial Endophytes Isolated from Plants in Natural Oil Seep Soils with Chronic Hydrocarbon Contamination
    Article Snippet: Paragraph title: Culture based analysis: endophyte isolation and identification ... The PCR reaction was as follows: 25 μL reactions with a final concentration of 0.5 mM of the forward primer and reverse primer, 1.5 mM MgCl2 , 200 mM of each dNTP, 2.5 units of HotStarTaq Plus DNA polymerase (Qiagen, Canada).

    Article Title: Effects of Upconversion Nanoparticles on Polymerase Chain Reaction
    Article Snippet: Top DNA polymerase, used in the AccuPower PCR PreMix, was a modified form of a recombinant DNA polymerase, originally isolated from Thermus thermophilus . .. The AmpliTaq Gold DNA Polymerase in the AmpliTaq Gold 360 Master Mix, and the HotStarTaq Plus DNA Polymerase in the HotStarTaq Plus Master Mix (Qiagen) were recombinant forms modified from the Thermus aquaticus (Taq ) DNA polymerase.

    Polymerase Chain Reaction:

    Article Title: Modulation of neurite outgrowth by activation of calcium-permeable kainate receptors expressed by rat nociceptive-like dorsal root ganglion neurons
    Article Snippet: A ‘master mix’ of the PCR reaction components was prepared before addition to individual PCR tubes to minimize pipetting errors, and all samples underwent PCR at the same time in the same experiment. .. Reaction mixture consisted of 1 μg of DRG or dorsal horn complementary DNA (cDNA), 10 pmol of each primer pair, 200 μM deoxynucleoside triphosphates (dNTPs), 50 mM KCl2, 10 mM Tris–HCl (pH 8.3), 1.5 mM MgCl2 , and 0.5 μl of HotStart Plus DNA polymerase (Qiagen).

    Article Title: Interplay between SIN3A and STAT3 Mediates Chromatin Conformational Changes and GFAP Expression during Cellular Differentiation
    Article Snippet: .. The PCRs were carried out with 25 ng bisulfite-treated DNA, 1 µl of HotStart Taq DNA polymerase (Qiagen), 10× supplied HotStart buffer, 4 µl of dNTP (25 mM), and 3 µl of PCR primers (5 µM). .. PCR conditions were 94°C for 15 min, then 40 cycles of 94°C for 30 s, 57°C for 30 s, and 72°C for 30 s. The PCR products were subjected to purification using the PCR cleanup kit (28160; Qiagen) following the manufacturer's instructions.

    Article Title: Effects of Upconversion Nanoparticles on Polymerase Chain Reaction
    Article Snippet: .. With the Qiagen HotStarTaq Plus DNA Polymerase in the Qiagen QuantiTect SYBR Green PCR Kit, the optimal concentrations of UCNPs for improving the PCR specificity were 0.5 µg/µL for the 40-nm-sized UCNPs, and 10× and 1× for the 70- and 250-nm-sized UCNPs, respectively, with a CT value of 13.44±0.07, 13.50±0.05, and 13.35±0.09, respectively (QuantiTect SYBR Green PCR Kit; 13.24±0.07 without UCNPs; ). .. PCR Amplification at Low Annealing Temperature At annealing temperatures ranging from 25°C to 60°C, specific PCR products were obtained, without nonspecific amplification, by addition of the 40 nm UCNP (1 µg/µL) to the PCR master mix ( ).

    Article Title: Valproic acid shows potent antitumor effect with alteration of DNA methylation in neuroblastoma
    Article Snippet: .. PCR Reactions were performed in triplicate in a 20 µl volume containing 1× buffer, 2U Hotstart Taq DNA polymerase (Qiagen), 250 nM of each primer, 1.5 µM SYTO-9, and 10 ng bisulfite-treated DNA template, with 2 mM final MgCl2 . ..

    Article Title: Alkaline-mediated differential interaction (AMDI): A simple automatable single-nucleotide polymorphism assay
    Article Snippet: All PCR reactions were performed with 100–120 ng of genomic DNA template in a final reaction volume of 15 μl. .. 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).

    Article Title: Structural studies of the catalytic core of the primate foamy virus (PFV-1) integrase
    Article Snippet: .. The PFV catalytic domain (amino acids 110–309) was amplified by PCR using oligonucleotide primers 5′-CATG CATATG GGTCCTATTCTAAGACCAGATAGG-3′ and 5′-CCGCTCGAGCTAAGAGGAGG CTGGAG GGGTGGATGG-3′ and HotStart HiFi Taq DNA polymerase (Qiagen). .. The PCR product was digested by Nde I and Bam HI (NEB) and cloned in pET-15b vector (Novagen) between Nde I and Xho I restriction sites.

    Article Title: Bacterial Endophytes Isolated from Plants in Natural Oil Seep Soils with Chronic Hydrocarbon Contamination
    Article Snippet: .. The PCR reaction was as follows: 25 μL reactions with a final concentration of 0.5 mM of the forward primer and reverse primer, 1.5 mM MgCl2 , 200 mM of each dNTP, 2.5 units of HotStarTaq Plus DNA polymerase (Qiagen, Canada). .. The PCR amplifications were carried out in a PTC-200 thermal cycler (MJ Research Inc.) with the following conditions: initial denaturing at 95°C for 5 min followed by 35 cycles of: denaturing at 95°C for 1 min, annealing at 56°C for 1 min and extension at 72°C for 1 min; final extension at 72°C for 10 min.

    Article Title: Effects of Upconversion Nanoparticles on Polymerase Chain Reaction
    Article Snippet: Paragraph title: 3. PCR Amplification ... The AmpliTaq Gold DNA Polymerase in the AmpliTaq Gold 360 Master Mix, and the HotStarTaq Plus DNA Polymerase in the HotStarTaq Plus Master Mix (Qiagen) were recombinant forms modified from the Thermus aquaticus (Taq ) DNA polymerase.

    Article Title: Pyrosequencing versus methylation-specific PCR for assessment of MGMT methylation in tumor and blood samples of glioblastoma patients
    Article Snippet: MGMT methylation assessment by MSP Bisulfite-converted DNA (2 μl) was amplified using specific primers (previously described for CpG sites 74–78 ) for methylated and unmethylated DNA independently, using HotStart®Plus DNA polymerase (Qiagen, Izasa, Spain) and following the manufacturer’s instructions. .. PCR reactions (15 μl) were analyzed on a 2% agarose gel stained with ethidium bromide or Syber Safe.

    Article Title: Gene expression in the developing mouse retina by EST sequencing and microarray analysis
    Article Snippet: .. Total RNA was reverse transcribed with Superscript reverse transcriptase (Life Technologies) at 42°C. cDNA equivalent to 40 ng of total RNA was used for PCR amplification with the Hotstart Taq DNA polymerase (Qiagen) in a volume of 20 µl. ..

    Purification:

    Article Title: Interplay between SIN3A and STAT3 Mediates Chromatin Conformational Changes and GFAP Expression during Cellular Differentiation
    Article Snippet: The PCRs were carried out with 25 ng bisulfite-treated DNA, 1 µl of HotStart Taq DNA polymerase (Qiagen), 10× supplied HotStart buffer, 4 µl of dNTP (25 mM), and 3 µl of PCR primers (5 µM). .. PCR conditions were 94°C for 15 min, then 40 cycles of 94°C for 30 s, 57°C for 30 s, and 72°C for 30 s. The PCR products were subjected to purification using the PCR cleanup kit (28160; Qiagen) following the manufacturer's instructions.

    Article Title: Tissue-specific transcriptomics, chromosomal localization, and phylogeny of chemosensory and odorant binding proteins from the red flour beetle Tribolium castaneum reveal subgroup specificities for olfaction or more general functions
    Article Snippet: Messenger RNA was purified with the Dynabeads purification kit (life technologies, Carlsbad, USA) and cDNA was synthesized using the Super-Script first-strand synthesis system (life technologies, Carlsbad, USA). .. Hotstart Taq DNA polymerase (Qiagen, Venlo, Netherlands) was used to amplify individual transcripts.

    Article Title: Bacterial Endophytes Isolated from Plants in Natural Oil Seep Soils with Chronic Hydrocarbon Contamination
    Article Snippet: Individual bacterial colonies were isolated and purified as they appeared. .. The PCR reaction was as follows: 25 μL reactions with a final concentration of 0.5 mM of the forward primer and reverse primer, 1.5 mM MgCl2 , 200 mM of each dNTP, 2.5 units of HotStarTaq Plus DNA polymerase (Qiagen, Canada).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Modulation of neurite outgrowth by activation of calcium-permeable kainate receptors expressed by rat nociceptive-like dorsal root ganglion neurons
    Article Snippet: Paragraph title: End-point RT-PCR ... Reaction mixture consisted of 1 μg of DRG or dorsal horn complementary DNA (cDNA), 10 pmol of each primer pair, 200 μM deoxynucleoside triphosphates (dNTPs), 50 mM KCl2, 10 mM Tris–HCl (pH 8.3), 1.5 mM MgCl2 , and 0.5 μl of HotStart Plus DNA polymerase (Qiagen).

    Gel Extraction:

    Article Title: Suberanilohydroxamic acid prevents TGF-β1-induced COX-2 repression in human lung fibroblasts post-transcriptionally by TIA-1 downregulation
    Article Snippet: COX -2 promoter region was amplified using 1 μM primers (primer sequences: forward 5′-GGTAGGAAATTTTATATTGGTGATT-3′ and reverse 5′-CTCACCTATATAACTAAACRCCA-3′), 2.5 mM MgCl2 , 1 μl HotStarTaq Plus DNA Polymerase (203601, Qiagen, Venlo, NL). .. DNA bands corresponding to COX -2 promoter were dissected under UV-light and the DNA was extracted using the QIAquick Gel Extraction Kit (28704, Qiagen) following the manufacturer's instructions.

    Plasmid Preparation:

    Article Title: Suberanilohydroxamic acid prevents TGF-β1-induced COX-2 repression in human lung fibroblasts post-transcriptionally by TIA-1 downregulation
    Article Snippet: COX -2 promoter region was amplified using 1 μM primers (primer sequences: forward 5′-GGTAGGAAATTTTATATTGGTGATT-3′ and reverse 5′-CTCACCTATATAACTAAACRCCA-3′), 2.5 mM MgCl2 , 1 μl HotStarTaq Plus DNA Polymerase (203601, Qiagen, Venlo, NL). .. DNA was then cloned using the pGEM®-T Easy Vector System II (A1380, Promega, Madison, WI, USA).

    Article Title: Interplay between SIN3A and STAT3 Mediates Chromatin Conformational Changes and GFAP Expression during Cellular Differentiation
    Article Snippet: The PCRs were carried out with 25 ng bisulfite-treated DNA, 1 µl of HotStart Taq DNA polymerase (Qiagen), 10× supplied HotStart buffer, 4 µl of dNTP (25 mM), and 3 µl of PCR primers (5 µM). .. The PCRs were carried out with 25 ng bisulfite-treated DNA, 1 µl of HotStart Taq DNA polymerase (Qiagen), 10× supplied HotStart buffer, 4 µl of dNTP (25 mM), and 3 µl of PCR primers (5 µM).

    Article Title: Tissue-specific transcriptomics, chromosomal localization, and phylogeny of chemosensory and odorant binding proteins from the red flour beetle Tribolium castaneum reveal subgroup specificities for olfaction or more general functions
    Article Snippet: Hotstart Taq DNA polymerase (Qiagen, Venlo, Netherlands) was used to amplify individual transcripts. .. Finally the products were cloned into PCR2.1 vector (life technologies, Carlsbad, USA) and verified by sequencing.

    Article Title: Structural studies of the catalytic core of the primate foamy virus (PFV-1) integrase
    Article Snippet: The PFV catalytic domain (amino acids 110–309) was amplified by PCR using oligonucleotide primers 5′-CATG CATATG GGTCCTATTCTAAGACCAGATAGG-3′ and 5′-CCGCTCGAGCTAAGAGGAGG CTGGAG GGGTGGATGG-3′ and HotStart HiFi Taq DNA polymerase (Qiagen). .. The PCR product was digested by Nde I and Bam HI (NEB) and cloned in pET-15b vector (Novagen) between Nde I and Xho I restriction sites.

    Real-time Polymerase Chain Reaction:

    Article Title: Effects of Upconversion Nanoparticles on Polymerase Chain Reaction
    Article Snippet: With the AccuPower PCR PreMix with Top Polymerase, the optimal concentrations of UCNPs for improving the PCR specificity were 1.0 µg/µL for the 40-nm-sized UCNPs, and 20×(1× = 2.4×105 particles/µL) and 15× for the 70- and 250-nm-sized UCNPs, respectively , and the CT value was 13.92±0.06, 14.18±0.36, and 14.67±0.05, respectively, in the real-time PCR (AccuPower GreenStar qPCR PreMix; 13.45±0.15 without UCNPs; ). .. With the Qiagen HotStarTaq Plus DNA Polymerase in the Qiagen QuantiTect SYBR Green PCR Kit, the optimal concentrations of UCNPs for improving the PCR specificity were 0.5 µg/µL for the 40-nm-sized UCNPs, and 10× and 1× for the 70- and 250-nm-sized UCNPs, respectively, with a CT value of 13.44±0.07, 13.50±0.05, and 13.35±0.09, respectively (QuantiTect SYBR Green PCR Kit; 13.24±0.07 without UCNPs; ).

    Article Title: Valproic acid shows potent antitumor effect with alteration of DNA methylation in neuroblastoma
    Article Snippet: PCR amplification and high resolution melting analysis (HRM) were performed on a 7500 Fast Real-Time PCR System. .. PCR Reactions were performed in triplicate in a 20 µl volume containing 1× buffer, 2U Hotstart Taq DNA polymerase (Qiagen), 250 nM of each primer, 1.5 µM SYTO-9, and 10 ng bisulfite-treated DNA template, with 2 mM final MgCl2 .

    Positron Emission Tomography:

    Article Title: Structural studies of the catalytic core of the primate foamy virus (PFV-1) integrase
    Article Snippet: The PFV catalytic domain (amino acids 110–309) was amplified by PCR using oligonucleotide primers 5′-CATG CATATG GGTCCTATTCTAAGACCAGATAGG-3′ and 5′-CCGCTCGAGCTAAGAGGAGG CTGGAG GGGTGGATGG-3′ and HotStart HiFi Taq DNA polymerase (Qiagen). .. The PCR product was digested by Nde I and Bam HI (NEB) and cloned in pET-15b vector (Novagen) between Nde I and Xho I restriction sites.

    Agarose Gel Electrophoresis:

    Article Title: Pyrosequencing versus methylation-specific PCR for assessment of MGMT methylation in tumor and blood samples of glioblastoma patients
    Article Snippet: MGMT methylation assessment by MSP Bisulfite-converted DNA (2 μl) was amplified using specific primers (previously described for CpG sites 74–78 ) for methylated and unmethylated DNA independently, using HotStart®Plus DNA polymerase (Qiagen, Izasa, Spain) and following the manufacturer’s instructions. .. PCR reactions (15 μl) were analyzed on a 2% agarose gel stained with ethidium bromide or Syber Safe.

    Evaporation:

    Article Title: Alkaline-mediated differential interaction (AMDI): A simple automatable single-nucleotide polymorphism assay
    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). .. Each well was sealed with a layer of mineral oil, and the plate was then sealed with an adhesive sealing sheet to prevent evaporation and cross contamination.

    Concentration Assay:

    Article Title: Bacterial Endophytes Isolated from Plants in Natural Oil Seep Soils with Chronic Hydrocarbon Contamination
    Article Snippet: .. The PCR reaction was as follows: 25 μL reactions with a final concentration of 0.5 mM of the forward primer and reverse primer, 1.5 mM MgCl2 , 200 mM of each dNTP, 2.5 units of HotStarTaq Plus DNA polymerase (Qiagen, Canada). .. The PCR amplifications were carried out in a PTC-200 thermal cycler (MJ Research Inc.) with the following conditions: initial denaturing at 95°C for 5 min followed by 35 cycles of: denaturing at 95°C for 1 min, annealing at 56°C for 1 min and extension at 72°C for 1 min; final extension at 72°C for 10 min.

    Staining:

    Article Title: Pyrosequencing versus methylation-specific PCR for assessment of MGMT methylation in tumor and blood samples of glioblastoma patients
    Article Snippet: MGMT methylation assessment by MSP Bisulfite-converted DNA (2 μl) was amplified using specific primers (previously described for CpG sites 74–78 ) for methylated and unmethylated DNA independently, using HotStart®Plus DNA polymerase (Qiagen, Izasa, Spain) and following the manufacturer’s instructions. .. PCR reactions (15 μl) were analyzed on a 2% agarose gel stained with ethidium bromide or Syber Safe.

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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 408 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 408 article reviews
    Price from $9.99 to $1999.99
    hotstar taq dna polymerase - by Bioz Stars, 2020-04
    99/100 stars
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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

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

    Journal: Genetics

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

    doi: 10.1534/genetics.113.161091

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

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

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