supertaq dna polymerase kit  (Thermo Fisher)


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    Structured Review

    Thermo Fisher supertaq dna polymerase kit
    Supertaq Dna Polymerase Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/supertaq dna polymerase kit/product/Thermo Fisher
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    supertaq dna polymerase kit - by Bioz Stars, 2020-03
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Genetic diversity in curtoviruses: a highly divergent strain of Beet mild curly top virus associated with an outbreak of curly top disease in pepper in Mexico
    Article Snippet: These overlapping primers were used in the PCR with proofreading DNA polymerase (Platinum Taq DNA polymerase High Fidelity, Invitrogen) to direct amplification of the genomic DNA of this curtovirus isolate. .. PCR-amplified DNA fragments of the expected size (~3.0 kb) were cloned using the pZero-Blunt cloning system (Invitrogen), and a recombinant plasmid (pMX-P24-1.0) with an ~3.0-kb insert was identified by restriction digestion analysis and then sequenced.

    Article Title: Generation of Aptamers from A Primer-Free Randomized ssDNA Library Using Magnetic-Assisted Rapid Aptamer Selection
    Article Snippet: Paragraph title: Primer-free aptamer cloning process ... The PCR reaction, which contained 1.25 U of DNA polymerase (Invitrogen), 0.1 mM of dNTPs, 0.5 mM of MgSO4 , and 0.5 nM of primers, was performed under the following conditions: 30 cycles at 40 seconds at 94 °C; 40 seconds at 60 °C, and 40 seconds at 72 °C.

    Article Title: Development of a Cellular Assay System To Study the Genome Replication of High- and Low-Risk Mucosal and Cutaneous Human Papillomaviruses ▿
    Article Snippet: HindIII linkers were added at the ends, and a subgenomic fragment of HPV-18 was cloned into the HindIII site in the pUC18 cloning vector, generating plasmid pUCeHPV18, which carries the early region of HPV-18. .. The E1 protein expression mutant pUCeHPV-18E1/BcuI was generated by digesting the HPV-18 subgenome construct with BcuI (Fermentas) and filling the BcuI site with a Klenow fragment of DNA polymerase (Fermentas) in the presence of deoxynucleoside triphosphates (dNTPs), followed by religation of the DNA.

    Amplification:

    Article Title: Genetic diversity in curtoviruses: a highly divergent strain of Beet mild curly top virus associated with an outbreak of curly top disease in pepper in Mexico
    Article Snippet: .. These overlapping primers were used in the PCR with proofreading DNA polymerase (Platinum Taq DNA polymerase High Fidelity, Invitrogen) to direct amplification of the genomic DNA of this curtovirus isolate. .. PCR-amplified DNA fragments of the expected size (~3.0 kb) were cloned using the pZero-Blunt cloning system (Invitrogen), and a recombinant plasmid (pMX-P24-1.0) with an ~3.0-kb insert was identified by restriction digestion analysis and then sequenced.

    Article Title: 27nt-RNAs guide histone variant deposition via ‘RNA-induced DNA replication interference’ and thus transmit parental genome partitioning in Stylonychia
    Article Snippet: .. Klenow reaction To analyse the effect of PIWI1/27nt-RNA in a linear DNA amplification reaction at ambient temperature, we performed a Klenow reaction using the large fragment of DNA polymerase (ThermoFisher) and pGEM-T-easy-PCNA as template upon manufacturers’ recommendations. .. For priming a modified T7 sequencing primer was used (Additional file : Table S1).

    Article Title: The TT8 Gene Encodes a Basic Helix-Loop-Helix Domain Protein Required for Expression of DFR and BAN Genes in Arabidopsis Siliques
    Article Snippet: .. The cDNA samples were diluted 10-fold, and 2 μL of dilution was amplified in a 50-μL PCR mixture containing DNA polymerase buffer (Gibco BRL), 2.5 mM MgCl2 , 200 μM of each deoxynucleotide triphosphate, 0.2 μM of each gene-specific primer, and 1 unit of Taq DNA polymerase (Gibco BRL). .. PCR was conducted for 18, 21, 24, 35, or 40 cycles with the following thermal profiles: 94°C for 30 sec (3 min for the first cycle), 60°C for 30 sec, and 72°C for 2 min 30 sec, with a 10-min terminal extension step at 72°C.

    Article Title: Generation of Aptamers from A Primer-Free Randomized ssDNA Library Using Magnetic-Assisted Rapid Aptamer Selection
    Article Snippet: Also, a partial sequence of ds-5′-stem-loop-CCCCC-TA-3′ ligated with yT & A-vector was also amplified. .. The PCR reaction, which contained 1.25 U of DNA polymerase (Invitrogen), 0.1 mM of dNTPs, 0.5 mM of MgSO4 , and 0.5 nM of primers, was performed under the following conditions: 30 cycles at 40 seconds at 94 °C; 40 seconds at 60 °C, and 40 seconds at 72 °C.

    DNA Ligation:

    Article Title: Generation of Aptamers from A Primer-Free Randomized ssDNA Library Using Magnetic-Assisted Rapid Aptamer Selection
    Article Snippet: The recovered ds-5′-S20 -stem-loop-CCCCC-cS20 -A-3′ and ds-5′-stem-loop-CCCCC-A-3′ were dispersed in 15 μl of ddH2 O and sub-cloned into a yT & A-vector (Yeastern Biotech, Taipei, Taiwan) through DNA ligation, which was performed in accordance to manufacturer instructions. .. The PCR reaction, which contained 1.25 U of DNA polymerase (Invitrogen), 0.1 mM of dNTPs, 0.5 mM of MgSO4 , and 0.5 nM of primers, was performed under the following conditions: 30 cycles at 40 seconds at 94 °C; 40 seconds at 60 °C, and 40 seconds at 72 °C.

    Quantitative RT-PCR:

    Article Title: The TT8 Gene Encodes a Basic Helix-Loop-Helix Domain Protein Required for Expression of DFR and BAN Genes in Arabidopsis Siliques
    Article Snippet: The cDNA samples were diluted 10-fold, and 2 μL of dilution was amplified in a 50-μL PCR mixture containing DNA polymerase buffer (Gibco BRL), 2.5 mM MgCl2 , 200 μM of each deoxynucleotide triphosphate, 0.2 μM of each gene-specific primer, and 1 unit of Taq DNA polymerase (Gibco BRL). .. For quantitative RT-PCR assays, reactions were performed with 18, 21, and 24 cycles to ensure that amplifications were within the linear range.

    Article Title: Stat3 and C/EBPβ synergize to induce miR-21 and miR-181b expression during sepsis
    Article Snippet: Paragraph title: Quantitative real-time PCR (RT-qPCR) ... The PCR reaction (25 μl) contained 5 μl ChIP DNA, 12.5 μl of 2x TaqMan real-time PCR Master Mix containing DNA polymerase and dNTPs (Applied Biosystem, Foster City, CA) and 100 nM of primer/probe mix (Integrated DNA Technologies).

    SYBR Green Assay:

    Article Title: Characterization of the transcripts and protein isoforms for cytoplasmic polyadenylation element binding protein-3 (CPEB3) in the mouse retina
    Article Snippet: .. Transcript-specific, exon-spanning primer sets were designed in house (table ) using Vector NTI; PCR master mix containing the DNA polymerase, dNTPs, optimized buffer, and SYBR Green was obtained from Applied Biosystems. .. PCR reactions were performed with an ABI 7300 real-time PCR system under the following conditions: 95°C, 10 min; 40 cycles of (95°C, 15 sec; 60°C, 1 min); 95°C, 15 sec; 60°C, 30 sec; 95°C, 15 sec.

    Article Title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
    Article Snippet: Reactions were incubated at 55°C for 5 min, followed by the addition of 25 μl 2× FailSafe PCR master mix (Epicentre), 1 μl FailSafe DNA polymerase (Epicentre), and 14 μl nuclease-free water (Ambion), and subsequent 5 min incubation at 72°C for nick translation. .. For each reaction, 2 μl was used as template for a real time PCR on a MiniOpticon (Bio-Rad) using 0.5 μl SYBR Green, 25 μl 2× Nextera PCR master mix, 1 μl Nextera PCR enzyme and nuclease-free water to 50 μl.

    Random Hexamer Labeling:

    Article Title: Massively parallel polymerase cloning and genome sequencing of single cells using nanoliter microwells
    Article Snippet: .. 10 U of DNA Polymerase I (Invitrogen, Carlsbad, CA) was added to the denatured amplicons along with 250 nanograms of unmodified random hexamer primer, 1 mM dNTPs, 1x Ampligase buffer (Epicentre, Madison, Wi), and 1x NEB buffer 2 (NEB, Cambridge, MA). ..

    Electrophoretic Mobility Shift Assay:

    Article Title: Quadruplex formation is necessary for stable PNA invasion into duplex DNA of BCL2 promoter region
    Article Snippet: .. For gel shift studies of plasmid fragments, after incubation with PNAs plasmids were cut by incubation with Fast Digest SpeI and PstI restriction enzymes (Fermentas, Hanover, MD, USA) for 20 min at 37°C followed by 3′-32 P labeling with [α-32 P]-dCTP (Perkin Elmer) by Klenow fragment of DNA polymerase I (Fermentas). .. After purification on G-25 microcolumns samples were run with gel electrophoresis.

    Expressing:

    Article Title: Development of a Cellular Assay System To Study the Genome Replication of High- and Low-Risk Mucosal and Cutaneous Human Papillomaviruses ▿
    Article Snippet: .. The E1 protein expression mutant pUCeHPV-18E1/BcuI was generated by digesting the HPV-18 subgenome construct with BcuI (Fermentas) and filling the BcuI site with a Klenow fragment of DNA polymerase (Fermentas) in the presence of deoxynucleoside triphosphates (dNTPs), followed by religation of the DNA. .. The E2 protein expression mutant pUCeHPV-18E2/StuI was generated in the subgenomic fragment of HPV-18 at the StuI site by insertion of a 22-bp oligonucleotide, 5′-TCCGGGTGATGCAAACCGGAGG-3′, with T4 ligase as described above (see Fig. 3H, line 1, for a schematic representation of the generated mutations).

    Modification:

    Article Title: 27nt-RNAs guide histone variant deposition via ‘RNA-induced DNA replication interference’ and thus transmit parental genome partitioning in Stylonychia
    Article Snippet: Klenow reaction To analyse the effect of PIWI1/27nt-RNA in a linear DNA amplification reaction at ambient temperature, we performed a Klenow reaction using the large fragment of DNA polymerase (ThermoFisher) and pGEM-T-easy-PCNA as template upon manufacturers’ recommendations. .. For priming a modified T7 sequencing primer was used (Additional file : Table S1).

    Countercurrent Chromatography:

    Article Title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
    Article Snippet: PCR-free library preparation Adaptor sequences (NoPCR1: 5'-AAT GAT ACG GCG ACC ACC GAG ATC TAC ACG CCT CCC TCG CGC CAT CAG AGA TGT GTA TAA GAG ACA G-3', and NoPCR2: 5'-CAA GCA GAA GAC GGC ATA CGA GAT CGG TCT GCC TTG CCA GCC CGC TCA GAG ATG TGT ATA AGA GAC AG-3') were designed to contain the original 'Nextera' adaptor sequences, but with an additional 5' overhang of either P1 or P2 on adaptor 1 or adaptor 2, respectively (i.e. sequences to make compatible with cluster PCR on Illumina flow-cell), thus eliminating the need to add them during a PCR step. .. Reactions were incubated at 55°C for 5 min, followed by the addition of 25 μl 2× FailSafe PCR master mix (Epicentre), 1 μl FailSafe DNA polymerase (Epicentre), and 14 μl nuclease-free water (Ambion), and subsequent 5 min incubation at 72°C for nick translation.

    Flow Cytometry:

    Article Title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
    Article Snippet: PCR-free library preparation Adaptor sequences (NoPCR1: 5'-AAT GAT ACG GCG ACC ACC GAG ATC TAC ACG CCT CCC TCG CGC CAT CAG AGA TGT GTA TAA GAG ACA G-3', and NoPCR2: 5'-CAA GCA GAA GAC GGC ATA CGA GAT CGG TCT GCC TTG CCA GCC CGC TCA GAG ATG TGT ATA AGA GAC AG-3') were designed to contain the original 'Nextera' adaptor sequences, but with an additional 5' overhang of either P1 or P2 on adaptor 1 or adaptor 2, respectively (i.e. sequences to make compatible with cluster PCR on Illumina flow-cell), thus eliminating the need to add them during a PCR step. .. Reactions were incubated at 55°C for 5 min, followed by the addition of 25 μl 2× FailSafe PCR master mix (Epicentre), 1 μl FailSafe DNA polymerase (Epicentre), and 14 μl nuclease-free water (Ambion), and subsequent 5 min incubation at 72°C for nick translation.

    Sequencing:

    Article Title: 27nt-RNAs guide histone variant deposition via ‘RNA-induced DNA replication interference’ and thus transmit parental genome partitioning in Stylonychia
    Article Snippet: Klenow reaction To analyse the effect of PIWI1/27nt-RNA in a linear DNA amplification reaction at ambient temperature, we performed a Klenow reaction using the large fragment of DNA polymerase (ThermoFisher) and pGEM-T-easy-PCNA as template upon manufacturers’ recommendations. .. For priming a modified T7 sequencing primer was used (Additional file : Table S1).

    Article Title: Generation of Aptamers from A Primer-Free Randomized ssDNA Library Using Magnetic-Assisted Rapid Aptamer Selection
    Article Snippet: Also, a partial sequence of ds-5′-stem-loop-CCCCC-TA-3′ ligated with yT & A-vector was also amplified. .. The PCR reaction, which contained 1.25 U of DNA polymerase (Invitrogen), 0.1 mM of dNTPs, 0.5 mM of MgSO4 , and 0.5 nM of primers, was performed under the following conditions: 30 cycles at 40 seconds at 94 °C; 40 seconds at 60 °C, and 40 seconds at 72 °C.

    Article Title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
    Article Snippet: The 5' phosphorylated reverse compliment of the 19 bp mosaic end (ME: 5'-Phos-CTG TCT CTT ATA CAC ATC T-3') sequence was hybridized to NoPCR1/2 by combining 5 μl of each NoPCR1 and NoPCR2 with 10 μl ME reverse complement all at 100 μM with 80 μl TE, followed by denaturation at 95°C for 5 min then slow cooling to room temperature for a final annealed adaptor concentration of 10 μM. .. Reactions were incubated at 55°C for 5 min, followed by the addition of 25 μl 2× FailSafe PCR master mix (Epicentre), 1 μl FailSafe DNA polymerase (Epicentre), and 14 μl nuclease-free water (Ambion), and subsequent 5 min incubation at 72°C for nick translation.

    Nick Translation:

    Article Title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
    Article Snippet: .. Reactions were incubated at 55°C for 5 min, followed by the addition of 25 μl 2× FailSafe PCR master mix (Epicentre), 1 μl FailSafe DNA polymerase (Epicentre), and 14 μl nuclease-free water (Ambion), and subsequent 5 min incubation at 72°C for nick translation. .. Tubes were then cleaned up using Qiaquick MinElute PCR purification columns (Qiagen), eluting in 12 μl buffer EB.

    Generated:

    Article Title: Expression of Kir7.1 and a Novel Kir7.1 Splice Variant in Native Human Retinal Pigment Epithelium
    Article Snippet: .. The PCR products were generated by adding DNA polymerase (Taq DNA polymerase; Gibco BRL Life Technologies, Gaithersburg, MD or SuperTaq-Plus™ ; Ambion, Austin, TX) and cycled 30 times for GAPDH or 35 times for Kir7.1, Kir7.1S, or rhodopsin (1 min at 94°C, 1 min at 54°C–64°C, 0.5 –1 min at 72°C), followed by a 7 min-extension at 72°C. ..

    Article Title: Development of a Cellular Assay System To Study the Genome Replication of High- and Low-Risk Mucosal and Cutaneous Human Papillomaviruses ▿
    Article Snippet: .. The E1 protein expression mutant pUCeHPV-18E1/BcuI was generated by digesting the HPV-18 subgenome construct with BcuI (Fermentas) and filling the BcuI site with a Klenow fragment of DNA polymerase (Fermentas) in the presence of deoxynucleoside triphosphates (dNTPs), followed by religation of the DNA. .. The E2 protein expression mutant pUCeHPV-18E2/StuI was generated in the subgenomic fragment of HPV-18 at the StuI site by insertion of a 22-bp oligonucleotide, 5′-TCCGGGTGATGCAAACCGGAGG-3′, with T4 ligase as described above (see Fig. 3H, line 1, for a schematic representation of the generated mutations).

    other:

    Article Title: A New Method of the Visualization of the Double-Stranded Mitochondrial and Nuclear DNA
    Article Snippet: DNA polymerase I (0.2 U/µl, 20 minutes, RT, Fermentas), a buffer for DNA polymerase I, 0.05 mM dATP, dGTP, dCTP and alternatively 0.05 mM Alexa 555-dUTP, biotin-dUTP or digoxigenin-dUTP; when biotin-dUTP and digoxigenin-dUTP were used, we also added dTTP or 5-bromo-2′-deoxyuridine-5′-triphosphate (BrdUTP).

    Article Title: Atomic Scissors: A New Method of Tracking the 5-Bromo-2?-Deoxyuridine-Labeled DNA In Situ
    Article Snippet: Enzymes used These enzymes and condition were used: Terminal deoxynucleotidyl transferase (TdT; 2 U/µl, 10 minutes, 37°C, Fermentas), buffer for TdT, 0.05 mM dATP, dGTP, dCTP and 0.05 mM Alexa Fluor® 555-aha-2′-deoxyuridine-5′-triphosphate (Alexa-dUTP); DNA polymerase I (0.2 U/µl, 10 minutes, RT, Fermentas), buffer for DNA polymerase I, 0.05 mM dATP, dGTP, dCTP and 0.05 mM Alexa-dUTP; Klenow fragment (0.2 U/µl, 10 minutes, RT, Fermentas), buffer for the Klenow fragment, 0.05 mM dATP, dGTP, dCTP and 0.05 mM Alexa-dUTP; Klenow fragment Exo- (0.2 U/µl, 10 minutes, RT, Fermentas), buffer for the Klenow fragment Exo-, 0.05 mM dATP, dGTP, dCTP and 0.05 mM Alexa-dUTP; Exonuclease III (1 U/µl, 30 minutes, RT, Fermentas), buffer for exonuclease III; Exonuclease λ (0.1 U/µl, 30 minutes, RT, Fermentas), buffer for exonuclease λ; Shrimp alkaline phosphomonoesterase (phosphatase; SAP; 1 U/µl, 20 minutes, 37°C, Fermentas), buffer for SAP.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: The TT8 Gene Encodes a Basic Helix-Loop-Helix Domain Protein Required for Expression of DFR and BAN Genes in Arabidopsis Siliques
    Article Snippet: For reverse transcription (RT)–PCR studies, 5 μg of DNA-free RNA extract was converted into first-strand cDNA by using the SuperScript preamplification system for first-strand cDNA synthesis (Gibco BRL) and oligo(dT)12–18 . .. The cDNA samples were diluted 10-fold, and 2 μL of dilution was amplified in a 50-μL PCR mixture containing DNA polymerase buffer (Gibco BRL), 2.5 mM MgCl2 , 200 μM of each deoxynucleotide triphosphate, 0.2 μM of each gene-specific primer, and 1 unit of Taq DNA polymerase (Gibco BRL).

    Article Title: Expression of Kir7.1 and a Novel Kir7.1 Splice Variant in Native Human Retinal Pigment Epithelium
    Article Snippet: Paragraph title: 2.4. Conventional RT-PCR Analysis ... The PCR products were generated by adding DNA polymerase (Taq DNA polymerase; Gibco BRL Life Technologies, Gaithersburg, MD or SuperTaq-Plus™ ; Ambion, Austin, TX) and cycled 30 times for GAPDH or 35 times for Kir7.1, Kir7.1S, or rhodopsin (1 min at 94°C, 1 min at 54°C–64°C, 0.5 –1 min at 72°C), followed by a 7 min-extension at 72°C.

    Recombinant:

    Article Title: Genetic diversity in curtoviruses: a highly divergent strain of Beet mild curly top virus associated with an outbreak of curly top disease in pepper in Mexico
    Article Snippet: These overlapping primers were used in the PCR with proofreading DNA polymerase (Platinum Taq DNA polymerase High Fidelity, Invitrogen) to direct amplification of the genomic DNA of this curtovirus isolate. .. PCR-amplified DNA fragments of the expected size (~3.0 kb) were cloned using the pZero-Blunt cloning system (Invitrogen), and a recombinant plasmid (pMX-P24-1.0) with an ~3.0-kb insert was identified by restriction digestion analysis and then sequenced.

    Nucleic Acid Electrophoresis:

    Article Title: Quadruplex formation is necessary for stable PNA invasion into duplex DNA of BCL2 promoter region
    Article Snippet: For gel shift studies of plasmid fragments, after incubation with PNAs plasmids were cut by incubation with Fast Digest SpeI and PstI restriction enzymes (Fermentas, Hanover, MD, USA) for 20 min at 37°C followed by 3′-32 P labeling with [α-32 P]-dCTP (Perkin Elmer) by Klenow fragment of DNA polymerase I (Fermentas). .. After purification on G-25 microcolumns samples were run with gel electrophoresis.

    Mutagenesis:

    Article Title: One-step genetic correction of hemoglobin E/beta-thalassemia patient-derived iPSCs by the CRISPR/Cas9 system
    Article Snippet: HbE mutation was detected by multiplex PCR. .. The PCR reaction consisted of 1.5 U of DNA polymerase (Platinum Taq; Invitrogen), 1× PCR buffer, 1.5 mM MgCl2 , 0.1 mM dNTPs, 0.4 μM HbE-Fc primer, 0.4 μM HbE-Rc primer, 0.5 μM HbE-Rn primer, 0.5 μM HbE-Fm primer and 2 μl of DNA sample in a total volume of 30 μl.

    Article Title: Development of a Cellular Assay System To Study the Genome Replication of High- and Low-Risk Mucosal and Cutaneous Human Papillomaviruses ▿
    Article Snippet: .. The E1 protein expression mutant pUCeHPV-18E1/BcuI was generated by digesting the HPV-18 subgenome construct with BcuI (Fermentas) and filling the BcuI site with a Klenow fragment of DNA polymerase (Fermentas) in the presence of deoxynucleoside triphosphates (dNTPs), followed by religation of the DNA. .. The E2 protein expression mutant pUCeHPV-18E2/StuI was generated in the subgenomic fragment of HPV-18 at the StuI site by insertion of a 22-bp oligonucleotide, 5′-TCCGGGTGATGCAAACCGGAGG-3′, with T4 ligase as described above (see Fig. 3H, line 1, for a schematic representation of the generated mutations).

    Isolation:

    Article Title: One-step genetic correction of hemoglobin E/beta-thalassemia patient-derived iPSCs by the CRISPR/Cas9 system
    Article Snippet: Genomic DNA was isolated using the Gentra® Puregene® Cell Kit (Qiagen) according to the manufacturer’s instructions. .. The PCR reaction consisted of 1.5 U of DNA polymerase (Platinum Taq; Invitrogen), 1× PCR buffer, 1.5 mM MgCl2 , 0.1 mM dNTPs, 0.4 μM HbE-Fc primer, 0.4 μM HbE-Rc primer, 0.5 μM HbE-Rn primer, 0.5 μM HbE-Fm primer and 2 μl of DNA sample in a total volume of 30 μl.

    Size-exclusion Chromatography:

    Article Title: The TT8 Gene Encodes a Basic Helix-Loop-Helix Domain Protein Required for Expression of DFR and BAN Genes in Arabidopsis Siliques
    Article Snippet: The cDNA samples were diluted 10-fold, and 2 μL of dilution was amplified in a 50-μL PCR mixture containing DNA polymerase buffer (Gibco BRL), 2.5 mM MgCl2 , 200 μM of each deoxynucleotide triphosphate, 0.2 μM of each gene-specific primer, and 1 unit of Taq DNA polymerase (Gibco BRL). .. PCR was conducted for 18, 21, 24, 35, or 40 cycles with the following thermal profiles: 94°C for 30 sec (3 min for the first cycle), 60°C for 30 sec, and 72°C for 2 min 30 sec, with a 10-min terminal extension step at 72°C.

    Article Title: Characterization of the transcripts and protein isoforms for cytoplasmic polyadenylation element binding protein-3 (CPEB3) in the mouse retina
    Article Snippet: PCR reactions were performed with an ABI 7300 real-time PCR system using the following conditions: 50°C, 2 min; 95°C, 10 min; 40 cycles of (95°C, 15 sec; 60°C, 1 min). .. Transcript-specific, exon-spanning primer sets were designed in house (table ) using Vector NTI; PCR master mix containing the DNA polymerase, dNTPs, optimized buffer, and SYBR Green was obtained from Applied Biosystems.

    Labeling:

    Article Title: Quadruplex formation is necessary for stable PNA invasion into duplex DNA of BCL2 promoter region
    Article Snippet: .. For gel shift studies of plasmid fragments, after incubation with PNAs plasmids were cut by incubation with Fast Digest SpeI and PstI restriction enzymes (Fermentas, Hanover, MD, USA) for 20 min at 37°C followed by 3′-32 P labeling with [α-32 P]-dCTP (Perkin Elmer) by Klenow fragment of DNA polymerase I (Fermentas). .. After purification on G-25 microcolumns samples were run with gel electrophoresis.

    Article Title: Stat3 and C/EBPβ synergize to induce miR-21 and miR-181b expression during sepsis
    Article Snippet: Quantitative real-time PCR (RT-qPCR) RT-qPCR was used to measure enrichment of miR-21 and miR-181b promoter sequences in the ChIPed DNA using primer and fluorescently labeled internal probe sequences specific to each promoter (Integrated DNA Technologies, Coralville, IA). .. The PCR reaction (25 μl) contained 5 μl ChIP DNA, 12.5 μl of 2x TaqMan real-time PCR Master Mix containing DNA polymerase and dNTPs (Applied Biosystem, Foster City, CA) and 100 nM of primer/probe mix (Integrated DNA Technologies).

    Purification:

    Article Title: Quadruplex formation is necessary for stable PNA invasion into duplex DNA of BCL2 promoter region
    Article Snippet: For gel shift studies of plasmid fragments, after incubation with PNAs plasmids were cut by incubation with Fast Digest SpeI and PstI restriction enzymes (Fermentas, Hanover, MD, USA) for 20 min at 37°C followed by 3′-32 P labeling with [α-32 P]-dCTP (Perkin Elmer) by Klenow fragment of DNA polymerase I (Fermentas). .. After purification on G-25 microcolumns samples were run with gel electrophoresis.

    Article Title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
    Article Snippet: Reactions were incubated at 55°C for 5 min, followed by the addition of 25 μl 2× FailSafe PCR master mix (Epicentre), 1 μl FailSafe DNA polymerase (Epicentre), and 14 μl nuclease-free water (Ambion), and subsequent 5 min incubation at 72°C for nick translation. .. Tubes were then cleaned up using Qiaquick MinElute PCR purification columns (Qiagen), eluting in 12 μl buffer EB.

    Polymerase Chain Reaction:

    Article Title: One-step genetic correction of hemoglobin E/beta-thalassemia patient-derived iPSCs by the CRISPR/Cas9 system
    Article Snippet: .. The PCR reaction consisted of 1.5 U of DNA polymerase (Platinum Taq; Invitrogen), 1× PCR buffer, 1.5 mM MgCl2 , 0.1 mM dNTPs, 0.4 μM HbE-Fc primer, 0.4 μM HbE-Rc primer, 0.5 μM HbE-Rn primer, 0.5 μM HbE-Fm primer and 2 μl of DNA sample in a total volume of 30 μl. .. The PCR was performed after an initial denaturation at 95 °C for 15 min followed by 30 cycles of denaturation (94 °C for 45 s), annealing (68 °C for 45 s) and extension (72 °C for 1 min), and a final extension step (72 °C for 7 min).

    Article Title: Genetic diversity in curtoviruses: a highly divergent strain of Beet mild curly top virus associated with an outbreak of curly top disease in pepper in Mexico
    Article Snippet: .. These overlapping primers were used in the PCR with proofreading DNA polymerase (Platinum Taq DNA polymerase High Fidelity, Invitrogen) to direct amplification of the genomic DNA of this curtovirus isolate. .. PCR-amplified DNA fragments of the expected size (~3.0 kb) were cloned using the pZero-Blunt cloning system (Invitrogen), and a recombinant plasmid (pMX-P24-1.0) with an ~3.0-kb insert was identified by restriction digestion analysis and then sequenced.

    Article Title: The TT8 Gene Encodes a Basic Helix-Loop-Helix Domain Protein Required for Expression of DFR and BAN Genes in Arabidopsis Siliques
    Article Snippet: .. The cDNA samples were diluted 10-fold, and 2 μL of dilution was amplified in a 50-μL PCR mixture containing DNA polymerase buffer (Gibco BRL), 2.5 mM MgCl2 , 200 μM of each deoxynucleotide triphosphate, 0.2 μM of each gene-specific primer, and 1 unit of Taq DNA polymerase (Gibco BRL). .. PCR was conducted for 18, 21, 24, 35, or 40 cycles with the following thermal profiles: 94°C for 30 sec (3 min for the first cycle), 60°C for 30 sec, and 72°C for 2 min 30 sec, with a 10-min terminal extension step at 72°C.

    Article Title: Generation of Aptamers from A Primer-Free Randomized ssDNA Library Using Magnetic-Assisted Rapid Aptamer Selection
    Article Snippet: .. The PCR reaction, which contained 1.25 U of DNA polymerase (Invitrogen), 0.1 mM of dNTPs, 0.5 mM of MgSO4 , and 0.5 nM of primers, was performed under the following conditions: 30 cycles at 40 seconds at 94 °C; 40 seconds at 60 °C, and 40 seconds at 72 °C. .. The PCR amplification increased the number of specific sequences in ligated ds-5′-S20 -stem-loop-CCCCC-cS20 -TA-3′, as well as in non-ligated ds-5′-stem-loop-CCCCC-TA-3′.

    Article Title: Expression of Kir7.1 and a Novel Kir7.1 Splice Variant in Native Human Retinal Pigment Epithelium
    Article Snippet: .. The PCR products were generated by adding DNA polymerase (Taq DNA polymerase; Gibco BRL Life Technologies, Gaithersburg, MD or SuperTaq-Plus™ ; Ambion, Austin, TX) and cycled 30 times for GAPDH or 35 times for Kir7.1, Kir7.1S, or rhodopsin (1 min at 94°C, 1 min at 54°C–64°C, 0.5 –1 min at 72°C), followed by a 7 min-extension at 72°C. ..

    Article Title: Characterization of the transcripts and protein isoforms for cytoplasmic polyadenylation element binding protein-3 (CPEB3) in the mouse retina
    Article Snippet: .. Transcript-specific, exon-spanning primer sets were designed in house (table ) using Vector NTI; PCR master mix containing the DNA polymerase, dNTPs, optimized buffer, and SYBR Green was obtained from Applied Biosystems. .. PCR reactions were performed with an ABI 7300 real-time PCR system under the following conditions: 95°C, 10 min; 40 cycles of (95°C, 15 sec; 60°C, 1 min); 95°C, 15 sec; 60°C, 30 sec; 95°C, 15 sec.

    Article Title: Stat3 and C/EBPβ synergize to induce miR-21 and miR-181b expression during sepsis
    Article Snippet: .. The PCR reaction (25 μl) contained 5 μl ChIP DNA, 12.5 μl of 2x TaqMan real-time PCR Master Mix containing DNA polymerase and dNTPs (Applied Biosystem, Foster City, CA) and 100 nM of primer/probe mix (Integrated DNA Technologies). .. The PCR conditions were: 2 min at 50°C, 10 min at 95°C, followed by 40 cycles with 15s at 95°C and 1 min at 60°C (combined annealing and extension), using the Bio-Rad CFX96 Real-Time System.

    Article Title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
    Article Snippet: .. Reactions were incubated at 55°C for 5 min, followed by the addition of 25 μl 2× FailSafe PCR master mix (Epicentre), 1 μl FailSafe DNA polymerase (Epicentre), and 14 μl nuclease-free water (Ambion), and subsequent 5 min incubation at 72°C for nick translation. .. Tubes were then cleaned up using Qiaquick MinElute PCR purification columns (Qiagen), eluting in 12 μl buffer EB.

    Microscopy:

    Article Title: The TT8 Gene Encodes a Basic Helix-Loop-Helix Domain Protein Required for Expression of DFR and BAN Genes in Arabidopsis Siliques
    Article Snippet: Cleared seeds were observed with a Microphot-FXA (Nikon, Tokyo, Japan) microscope equipped with Nomarski differential interference contrast optics. .. The cDNA samples were diluted 10-fold, and 2 μL of dilution was amplified in a 50-μL PCR mixture containing DNA polymerase buffer (Gibco BRL), 2.5 mM MgCl2 , 200 μM of each deoxynucleotide triphosphate, 0.2 μM of each gene-specific primer, and 1 unit of Taq DNA polymerase (Gibco BRL).

    IA:

    Article Title: Stat3 and C/EBPβ synergize to induce miR-21 and miR-181b expression during sepsis
    Article Snippet: Quantitative real-time PCR (RT-qPCR) RT-qPCR was used to measure enrichment of miR-21 and miR-181b promoter sequences in the ChIPed DNA using primer and fluorescently labeled internal probe sequences specific to each promoter (Integrated DNA Technologies, Coralville, IA). .. The PCR reaction (25 μl) contained 5 μl ChIP DNA, 12.5 μl of 2x TaqMan real-time PCR Master Mix containing DNA polymerase and dNTPs (Applied Biosystem, Foster City, CA) and 100 nM of primer/probe mix (Integrated DNA Technologies).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
    Article Snippet: PCR-free library preparation Adaptor sequences (NoPCR1: 5'-AAT GAT ACG GCG ACC ACC GAG ATC TAC ACG CCT CCC TCG CGC CAT CAG AGA TGT GTA TAA GAG ACA G-3', and NoPCR2: 5'-CAA GCA GAA GAC GGC ATA CGA GAT CGG TCT GCC TTG CCA GCC CGC TCA GAG ATG TGT ATA AGA GAC AG-3') were designed to contain the original 'Nextera' adaptor sequences, but with an additional 5' overhang of either P1 or P2 on adaptor 1 or adaptor 2, respectively (i.e. sequences to make compatible with cluster PCR on Illumina flow-cell), thus eliminating the need to add them during a PCR step. .. Reactions were incubated at 55°C for 5 min, followed by the addition of 25 μl 2× FailSafe PCR master mix (Epicentre), 1 μl FailSafe DNA polymerase (Epicentre), and 14 μl nuclease-free water (Ambion), and subsequent 5 min incubation at 72°C for nick translation.

    Chromatin Immunoprecipitation:

    Article Title: Stat3 and C/EBPβ synergize to induce miR-21 and miR-181b expression during sepsis
    Article Snippet: .. The PCR reaction (25 μl) contained 5 μl ChIP DNA, 12.5 μl of 2x TaqMan real-time PCR Master Mix containing DNA polymerase and dNTPs (Applied Biosystem, Foster City, CA) and 100 nM of primer/probe mix (Integrated DNA Technologies). .. The PCR conditions were: 2 min at 50°C, 10 min at 95°C, followed by 40 cycles with 15s at 95°C and 1 min at 60°C (combined annealing and extension), using the Bio-Rad CFX96 Real-Time System.

    Plasmid Preparation:

    Article Title: Genetic diversity in curtoviruses: a highly divergent strain of Beet mild curly top virus associated with an outbreak of curly top disease in pepper in Mexico
    Article Snippet: These overlapping primers were used in the PCR with proofreading DNA polymerase (Platinum Taq DNA polymerase High Fidelity, Invitrogen) to direct amplification of the genomic DNA of this curtovirus isolate. .. PCR-amplified DNA fragments of the expected size (~3.0 kb) were cloned using the pZero-Blunt cloning system (Invitrogen), and a recombinant plasmid (pMX-P24-1.0) with an ~3.0-kb insert was identified by restriction digestion analysis and then sequenced.

    Article Title: Quadruplex formation is necessary for stable PNA invasion into duplex DNA of BCL2 promoter region
    Article Snippet: .. For gel shift studies of plasmid fragments, after incubation with PNAs plasmids were cut by incubation with Fast Digest SpeI and PstI restriction enzymes (Fermentas, Hanover, MD, USA) for 20 min at 37°C followed by 3′-32 P labeling with [α-32 P]-dCTP (Perkin Elmer) by Klenow fragment of DNA polymerase I (Fermentas). .. After purification on G-25 microcolumns samples were run with gel electrophoresis.

    Article Title: Characterization of the transcripts and protein isoforms for cytoplasmic polyadenylation element binding protein-3 (CPEB3) in the mouse retina
    Article Snippet: .. Transcript-specific, exon-spanning primer sets were designed in house (table ) using Vector NTI; PCR master mix containing the DNA polymerase, dNTPs, optimized buffer, and SYBR Green was obtained from Applied Biosystems. .. PCR reactions were performed with an ABI 7300 real-time PCR system under the following conditions: 95°C, 10 min; 40 cycles of (95°C, 15 sec; 60°C, 1 min); 95°C, 15 sec; 60°C, 30 sec; 95°C, 15 sec.

    Article Title: Development of a Cellular Assay System To Study the Genome Replication of High- and Low-Risk Mucosal and Cutaneous Human Papillomaviruses ▿
    Article Snippet: The E1 protein expression mutant pUCeHPV-18E1/OliI was generated by digestion of the subgenomic plasmid with OliI (a single site in the E1 ORF), and a 22-bp oligonucleotide, 5′-TCCGGGTGATGCAAACCGGAGG-3′, was inserted into the OliI site with T4 ligase. .. The E1 protein expression mutant pUCeHPV-18E1/BcuI was generated by digesting the HPV-18 subgenome construct with BcuI (Fermentas) and filling the BcuI site with a Klenow fragment of DNA polymerase (Fermentas) in the presence of deoxynucleoside triphosphates (dNTPs), followed by religation of the DNA.

    Software:

    Article Title: Genetic diversity in curtoviruses: a highly divergent strain of Beet mild curly top virus associated with an outbreak of curly top disease in pepper in Mexico
    Article Snippet: These overlapping primers were used in the PCR with proofreading DNA polymerase (Platinum Taq DNA polymerase High Fidelity, Invitrogen) to direct amplification of the genomic DNA of this curtovirus isolate. .. DNA sequences were analyzed and assembled with Vector NTI software (Version 10; Invitrogen).

    Article Title: Characterization of the transcripts and protein isoforms for cytoplasmic polyadenylation element binding protein-3 (CPEB3) in the mouse retina
    Article Snippet: Data were analyzed with the aid of SDS2.1 software (Applied Biosystems). .. Transcript-specific, exon-spanning primer sets were designed in house (table ) using Vector NTI; PCR master mix containing the DNA polymerase, dNTPs, optimized buffer, and SYBR Green was obtained from Applied Biosystems.

    Real-time Polymerase Chain Reaction:

    Article Title: 27nt-RNAs guide histone variant deposition via ‘RNA-induced DNA replication interference’ and thus transmit parental genome partitioning in Stylonychia
    Article Snippet: Klenow reaction To analyse the effect of PIWI1/27nt-RNA in a linear DNA amplification reaction at ambient temperature, we performed a Klenow reaction using the large fragment of DNA polymerase (ThermoFisher) and pGEM-T-easy-PCNA as template upon manufacturers’ recommendations. .. The amount of polymerized PCNA sequence was successively analysed using qPCR as described above.

    Article Title: Characterization of the transcripts and protein isoforms for cytoplasmic polyadenylation element binding protein-3 (CPEB3) in the mouse retina
    Article Snippet: Paragraph title: Real-time PCR ... Transcript-specific, exon-spanning primer sets were designed in house (table ) using Vector NTI; PCR master mix containing the DNA polymerase, dNTPs, optimized buffer, and SYBR Green was obtained from Applied Biosystems.

    Article Title: Stat3 and C/EBPβ synergize to induce miR-21 and miR-181b expression during sepsis
    Article Snippet: .. The PCR reaction (25 μl) contained 5 μl ChIP DNA, 12.5 μl of 2x TaqMan real-time PCR Master Mix containing DNA polymerase and dNTPs (Applied Biosystem, Foster City, CA) and 100 nM of primer/probe mix (Integrated DNA Technologies). .. The PCR conditions were: 2 min at 50°C, 10 min at 95°C, followed by 40 cycles with 15s at 95°C and 1 min at 60°C (combined annealing and extension), using the Bio-Rad CFX96 Real-Time System.

    Article Title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
    Article Snippet: Reactions were incubated at 55°C for 5 min, followed by the addition of 25 μl 2× FailSafe PCR master mix (Epicentre), 1 μl FailSafe DNA polymerase (Epicentre), and 14 μl nuclease-free water (Ambion), and subsequent 5 min incubation at 72°C for nick translation. .. For each reaction, 2 μl was used as template for a real time PCR on a MiniOpticon (Bio-Rad) using 0.5 μl SYBR Green, 25 μl 2× Nextera PCR master mix, 1 μl Nextera PCR enzyme and nuclease-free water to 50 μl.

    Multiplex Assay:

    Article Title: One-step genetic correction of hemoglobin E/beta-thalassemia patient-derived iPSCs by the CRISPR/Cas9 system
    Article Snippet: Paragraph title: Multiplex PCR analysis for hemoglobin E ... The PCR reaction consisted of 1.5 U of DNA polymerase (Platinum Taq; Invitrogen), 1× PCR buffer, 1.5 mM MgCl2 , 0.1 mM dNTPs, 0.4 μM HbE-Fc primer, 0.4 μM HbE-Rc primer, 0.5 μM HbE-Rn primer, 0.5 μM HbE-Fm primer and 2 μl of DNA sample in a total volume of 30 μl.

    Agarose Gel Electrophoresis:

    Article Title: The TT8 Gene Encodes a Basic Helix-Loop-Helix Domain Protein Required for Expression of DFR and BAN Genes in Arabidopsis Siliques
    Article Snippet: The cDNA samples were diluted 10-fold, and 2 μL of dilution was amplified in a 50-μL PCR mixture containing DNA polymerase buffer (Gibco BRL), 2.5 mM MgCl2 , 200 μM of each deoxynucleotide triphosphate, 0.2 μM of each gene-specific primer, and 1 unit of Taq DNA polymerase (Gibco BRL). .. The PCR products were size-separated on a 1% (w/v) agarose gel, blotted onto positively charged nylon membrane (GeneScreen Plus; Du Pont), and hybridized with primed 32 P-labeled fragments.

    Article Title: Generation of Aptamers from A Primer-Free Randomized ssDNA Library Using Magnetic-Assisted Rapid Aptamer Selection
    Article Snippet: The PCR reaction, which contained 1.25 U of DNA polymerase (Invitrogen), 0.1 mM of dNTPs, 0.5 mM of MgSO4 , and 0.5 nM of primers, was performed under the following conditions: 30 cycles at 40 seconds at 94 °C; 40 seconds at 60 °C, and 40 seconds at 72 °C. .. The PCR product was mixed with 6X DNA loading dye (0.25% bromophenol blue, 0.25% xylene cyanol, and 15% Ficoll) and the sample was run using a 3% agarose gel.

    Electrophoresis:

    Article Title: Generation of Aptamers from A Primer-Free Randomized ssDNA Library Using Magnetic-Assisted Rapid Aptamer Selection
    Article Snippet: The PCR reaction, which contained 1.25 U of DNA polymerase (Invitrogen), 0.1 mM of dNTPs, 0.5 mM of MgSO4 , and 0.5 nM of primers, was performed under the following conditions: 30 cycles at 40 seconds at 94 °C; 40 seconds at 60 °C, and 40 seconds at 72 °C. .. The electrophoresis result was analyzed by Alpha Imager EC (Alpha Innotech Corporation, San Leandro, CA, USA) and the full-length gel is presented in .

    Ethanol Precipitation:

    Article Title: Massively parallel polymerase cloning and genome sequencing of single cells using nanoliter microwells
    Article Snippet: 10 U of DNA Polymerase I (Invitrogen, Carlsbad, CA) was added to the denatured amplicons along with 250 nanograms of unmodified random hexamer primer, 1 mM dNTPs, 1x Ampligase buffer (Epicentre, Madison, Wi), and 1x NEB buffer 2 (NEB, Cambridge, MA). .. The reaction was cleaned using standard ethanol precipitation and eluted in 4 μL water.

    Incubation:

    Article Title: Homogeneous antibody-based proximity extension assays provide sensitive and specific detection of low-abundant proteins in human blood
    Article Snippet: .. After a 5-min incubation at 37°C, a 20 µl extension mix containing 66.8 mM Tris–HCl, 16.8 mM ammonium sulfate, 1 mM dithiothreitol, 33 mM magnesium chloride, 62.5 U/ml T4 DNA Polymerase [or 62.5 U/ml Klenow fragment exo(−), 125 U/ml Klenow fragment, 125 U/ml DNA Polymerase I (Fermentas), 250 U/ml Exonuclease I (New England Biolabs)] was added. ..

    Article Title: Massively parallel polymerase cloning and genome sequencing of single cells using nanoliter microwells
    Article Snippet: Low-input library construction 1.5 μL of ALS buffer was added to the extracted amplicons to denature the DNA followed by a 3-minute incubation at room temperature. .. 10 U of DNA Polymerase I (Invitrogen, Carlsbad, CA) was added to the denatured amplicons along with 250 nanograms of unmodified random hexamer primer, 1 mM dNTPs, 1x Ampligase buffer (Epicentre, Madison, Wi), and 1x NEB buffer 2 (NEB, Cambridge, MA).

    Article Title: Quadruplex formation is necessary for stable PNA invasion into duplex DNA of BCL2 promoter region
    Article Snippet: .. For gel shift studies of plasmid fragments, after incubation with PNAs plasmids were cut by incubation with Fast Digest SpeI and PstI restriction enzymes (Fermentas, Hanover, MD, USA) for 20 min at 37°C followed by 3′-32 P labeling with [α-32 P]-dCTP (Perkin Elmer) by Klenow fragment of DNA polymerase I (Fermentas). .. After purification on G-25 microcolumns samples were run with gel electrophoresis.

    Article Title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
    Article Snippet: .. Reactions were incubated at 55°C for 5 min, followed by the addition of 25 μl 2× FailSafe PCR master mix (Epicentre), 1 μl FailSafe DNA polymerase (Epicentre), and 14 μl nuclease-free water (Ambion), and subsequent 5 min incubation at 72°C for nick translation. .. Tubes were then cleaned up using Qiaquick MinElute PCR purification columns (Qiagen), eluting in 12 μl buffer EB.

    Produced:

    Article Title: Development of a Cellular Assay System To Study the Genome Replication of High- and Low-Risk Mucosal and Cutaneous Human Papillomaviruses ▿
    Article Snippet: The E1 protein expression mutant pUCeHPV-18E1/BcuI was generated by digesting the HPV-18 subgenome construct with BcuI (Fermentas) and filling the BcuI site with a Klenow fragment of DNA polymerase (Fermentas) in the presence of deoxynucleoside triphosphates (dNTPs), followed by religation of the DNA. .. The defects of the generated viral mutants in a transient-replication assay in U2OS cells and complementation of these defects in the viral genome by the respective proteins produced by the cotransfected expression vectors were demonstrated.

    Concentration Assay:

    Article Title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition
    Article Snippet: The 5' phosphorylated reverse compliment of the 19 bp mosaic end (ME: 5'-Phos-CTG TCT CTT ATA CAC ATC T-3') sequence was hybridized to NoPCR1/2 by combining 5 μl of each NoPCR1 and NoPCR2 with 10 μl ME reverse complement all at 100 μM with 80 μl TE, followed by denaturation at 95°C for 5 min then slow cooling to room temperature for a final annealed adaptor concentration of 10 μM. .. Reactions were incubated at 55°C for 5 min, followed by the addition of 25 μl 2× FailSafe PCR master mix (Epicentre), 1 μl FailSafe DNA polymerase (Epicentre), and 14 μl nuclease-free water (Ambion), and subsequent 5 min incubation at 72°C for nick translation.

    Construct:

    Article Title: Development of a Cellular Assay System To Study the Genome Replication of High- and Low-Risk Mucosal and Cutaneous Human Papillomaviruses ▿
    Article Snippet: .. The E1 protein expression mutant pUCeHPV-18E1/BcuI was generated by digesting the HPV-18 subgenome construct with BcuI (Fermentas) and filling the BcuI site with a Klenow fragment of DNA polymerase (Fermentas) in the presence of deoxynucleoside triphosphates (dNTPs), followed by religation of the DNA. .. The E2 protein expression mutant pUCeHPV-18E2/StuI was generated in the subgenomic fragment of HPV-18 at the StuI site by insertion of a 22-bp oligonucleotide, 5′-TCCGGGTGATGCAAACCGGAGG-3′, with T4 ligase as described above (see Fig. 3H, line 1, for a schematic representation of the generated mutations).

    DNA Purification:

    Article Title: Generation of Aptamers from A Primer-Free Randomized ssDNA Library Using Magnetic-Assisted Rapid Aptamer Selection
    Article Snippet: A DNA purification process was performed to remove enzyme and reaction buffer, and to recover ds-5′-S20 -stem-loop-CCCCC-cS20 -A-3′ and ds-5′-stem-loop-CCCCC-A-3′. .. The PCR reaction, which contained 1.25 U of DNA polymerase (Invitrogen), 0.1 mM of dNTPs, 0.5 mM of MgSO4 , and 0.5 nM of primers, was performed under the following conditions: 30 cycles at 40 seconds at 94 °C; 40 seconds at 60 °C, and 40 seconds at 72 °C.

    Variant Assay:

    Article Title: Expression of Kir7.1 and a Novel Kir7.1 Splice Variant in Native Human Retinal Pigment Epithelium
    Article Snippet: PCR was performed with gene- or splice variant-specific primer sets ( ). .. The PCR products were generated by adding DNA polymerase (Taq DNA polymerase; Gibco BRL Life Technologies, Gaithersburg, MD or SuperTaq-Plus™ ; Ambion, Austin, TX) and cycled 30 times for GAPDH or 35 times for Kir7.1, Kir7.1S, or rhodopsin (1 min at 94°C, 1 min at 54°C–64°C, 0.5 –1 min at 72°C), followed by a 7 min-extension at 72°C.

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    Thermo Fisher gene exp rna polymerase ii gm03758725 g1
    ( A–C ) Correlation analysis of the global miRNAs expression between different amounts of total <t>RNA</t> input (1, 3, and 5 μg) from FFPE samples.
    Gene Exp Rna Polymerase Ii Gm03758725 G1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp rna polymerase ii gm03758725 g1/product/Thermo Fisher
    Average 91 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    gene exp rna polymerase ii gm03758725 g1 - by Bioz Stars, 2020-03
    91/100 stars
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    99
    Thermo Fisher supertaq dna polymerase
    ( A–C ) Correlation analysis of the global miRNAs expression between different amounts of total <t>RNA</t> input (1, 3, and 5 μg) from FFPE samples.
    Supertaq Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/supertaq dna polymerase/product/Thermo Fisher
    Average 99 stars, based on 12 article reviews
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    supertaq dna polymerase - by Bioz Stars, 2020-03
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    ( A–C ) Correlation analysis of the global miRNAs expression between different amounts of total RNA input (1, 3, and 5 μg) from FFPE samples.

    Journal:

    Article Title: Systematic analysis of microRNA expression of RNA extracted from fresh frozen and formalin-fixed paraffin-embedded samples

    doi: 10.1261/rna.642907

    Figure Lengend Snippet: ( A–C ) Correlation analysis of the global miRNAs expression between different amounts of total RNA input (1, 3, and 5 μg) from FFPE samples.

    Article Snippet: The PCR master mix containing mir Vana 5× PCR buffer (with SYBR Green I), 50× ROX, SuperTaq Polymerase, mir Vana PCR primers, and RT products were processed as follows: 95°C for 3 min and then 40 cycles of 95°C for 15 sec and 60°C for 35 sec ( n = 3).

    Techniques: Expressing, Formalin-fixed Paraffin-Embedded

    Correlation analysis of the messenger RNA expression between fresh frozen and FFPE samples. ( A ) Global mRNAs expression comparison. ( B ) Overlapping mRNAs expression comparison.

    Journal:

    Article Title: Systematic analysis of microRNA expression of RNA extracted from fresh frozen and formalin-fixed paraffin-embedded samples

    doi: 10.1261/rna.642907

    Figure Lengend Snippet: Correlation analysis of the messenger RNA expression between fresh frozen and FFPE samples. ( A ) Global mRNAs expression comparison. ( B ) Overlapping mRNAs expression comparison.

    Article Snippet: The PCR master mix containing mir Vana 5× PCR buffer (with SYBR Green I), 50× ROX, SuperTaq Polymerase, mir Vana PCR primers, and RT products were processed as follows: 95°C for 3 min and then 40 cycles of 95°C for 15 sec and 60°C for 35 sec ( n = 3).

    Techniques: RNA Expression, Formalin-fixed Paraffin-Embedded, Expressing

    Effect of formalin fixation on miRNA expression. ( A ) Representative RNA agarose gel image (lane 1 , RNA ladder; lane 2 , fresh frozen sample; lane 3 , FFPE sample). ( B ) The expression of hsa - miR-16 and RUN6B was analyzed using real-time QRT-PCR analysis

    Journal:

    Article Title: Systematic analysis of microRNA expression of RNA extracted from fresh frozen and formalin-fixed paraffin-embedded samples

    doi: 10.1261/rna.642907

    Figure Lengend Snippet: Effect of formalin fixation on miRNA expression. ( A ) Representative RNA agarose gel image (lane 1 , RNA ladder; lane 2 , fresh frozen sample; lane 3 , FFPE sample). ( B ) The expression of hsa - miR-16 and RUN6B was analyzed using real-time QRT-PCR analysis

    Article Snippet: The PCR master mix containing mir Vana 5× PCR buffer (with SYBR Green I), 50× ROX, SuperTaq Polymerase, mir Vana PCR primers, and RT products were processed as follows: 95°C for 3 min and then 40 cycles of 95°C for 15 sec and 60°C for 35 sec ( n = 3).

    Techniques: Expressing, Agarose Gel Electrophoresis, Formalin-fixed Paraffin-Embedded, Quantitative RT-PCR

    Correlation analysis of the global miRNAs expression between fresh frozen and FFPE samples. ( A–C ) The comparisons of different input RNA amounts (1, 3, and 5 μg) of FFPE samples versus 1 μg fresh frozen sample.

    Journal:

    Article Title: Systematic analysis of microRNA expression of RNA extracted from fresh frozen and formalin-fixed paraffin-embedded samples

    doi: 10.1261/rna.642907

    Figure Lengend Snippet: Correlation analysis of the global miRNAs expression between fresh frozen and FFPE samples. ( A–C ) The comparisons of different input RNA amounts (1, 3, and 5 μg) of FFPE samples versus 1 μg fresh frozen sample.

    Article Snippet: The PCR master mix containing mir Vana 5× PCR buffer (with SYBR Green I), 50× ROX, SuperTaq Polymerase, mir Vana PCR primers, and RT products were processed as follows: 95°C for 3 min and then 40 cycles of 95°C for 15 sec and 60°C for 35 sec ( n = 3).

    Techniques: Expressing, Formalin-fixed Paraffin-Embedded