dna polymerase  (Thermo Fisher)


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    Structured Review

    Thermo Fisher dna polymerase
    <t>PCR</t> of human genomic <t>DNA</t>
    Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 403 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna polymerase/product/Thermo Fisher
    Average 97 stars, based on 403 article reviews
    Price from $9.99 to $1999.99
    dna polymerase - by Bioz Stars, 2020-02
    97/100 stars

    Images

    1) Product Images from "Expression of Kir7.1 and a Novel Kir7.1 Splice Variant in Native Human Retinal Pigment Epithelium"

    Article Title: Expression of Kir7.1 and a Novel Kir7.1 Splice Variant in Native Human Retinal Pigment Epithelium

    Journal:

    doi: 10.1016/j.exer.2007.09.011

    PCR of human genomic DNA
    Figure Legend Snippet: PCR of human genomic DNA

    Techniques Used: Polymerase Chain Reaction

    Related Articles

    Clone Assay:

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Paragraph title: Generation of gene cluster DNA fragments and yeast assembly cloning in the yTREX vector ... Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Amplification:

    Article Title: Correlation of intestinal disaccharidase activities with the C/T-13910 variant and age
    Article Snippet: The DNA fragment spanning the C/T-13910 variant was amplified using a biotinylated and an unbiotinylated primer (primers available upon request). .. Typically, 50 μL of the minisequencing reaction mixture contained 10 pmoles of the minisequencing primers for C/T-13910 and 0.1 μL of either 3H-dCTP corresponding to the lactase non-persistence allele (Amersham, UK) or 3H-dTTP corresponding to the lactase persistence allele, and 0.05 units of DNA polymerase (Dynazyme II, Finnzymes).

    Article Title: Single-amplicon MSH2 A636P Mutation Testing in Ashkenazi Jewish Patients With Colorectal Cancer
    Article Snippet: .. The PCR amplification was performed in a final volume of 50 μL, containing 50 ng of DNA, 2.5 mM dNTPs, 15 pmoles of each primer, and 1.25 U of DNA polymerase (AmpliTaq Gold, Applied Biosystems) in Buffer II. .. The PCR amplification consisted of 10 minutes at 95°C, followed by 35 cycles of 30 seconds at 94°C, 30 seconds at 55°C, and 45 seconds at 72°C; a final extension of 7 minutes ended the amplification reaction.

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: Paragraph title: Sample procurement and RNA amplification: ... Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr.

    Article Title: The C/C−13910 genotype of adult-type hypolactasia is associated with an increased risk of colorectal cancer in the Finnish population
    Article Snippet: The DNA fragment spanning the C/T−13910 variant was amplified using one biotinylated (5′-CCT CGT TAA TAC CCA CTG ACC TA-3′) primer and one unbiotinylated (5′-GTC ACT TTG ATA TGA TGA GAG CA-3′) primer. .. The minisequencing reaction contained 10 pmol of the minisequencing primer for C/T−13910 (5′-GGC AAT ACA GAT AAG ATA ATG TAG-3′) and 0.1 μl of either H-dCTP or H-dTTP (Amersham Biosciences, Little Chalfont, Buckinghamshire, UK), and 0.05 U of DNA polymerase (Dynazyme II, Finnzymes, Espoo, Finland).

    Article Title: Gene Conversion Between Direct Noncoding Repeats Promotes Genetic and Phenotypic Diversity at a Regulatory Locus of Zea mays (L.)
    Article Snippet: Paragraph title: Amplification and sequencing of the p1 noncoding regions: ... The PCR reactions were performed using enhanced DNA polymerase, Elongase (Invitrogen, Carlsbad, CA) with ∼1 min of extension time for every 1-kb fragment size.

    Article Title: An RNA interference model of RPS19 deficiency in Diamond-Blackfan anemia recapitulates defective hematopoiesis and rescue by dexamethasone: identification of dexamethasone-responsive genes by microarray
    Article Snippet: Briefly, first-round amplification was carried out with oligo(dT)-T7 primers and Superscript II (Invitrogen) in the presence of the single-stranded nucleic acid binding protein T4gp32 (USB) for one hour at 4°C. .. After second-strand synthesis in the presence of DNA polymerase, Escherichia coli RNase H, and E coli DNA ligase (Invitrogen, Cleveland, OH), the DNA was polished with T4 DNA polymerase and purified by phenol/chloroform extraction followed by chromatography on a BioGel P-6 MicroSpin Column (BioRad, Hercules, CA).

    Synthesized:

    Article Title: An RNA interference model of RPS19 deficiency in Diamond-Blackfan anemia recapitulates defective hematopoiesis and rescue by dexamethasone: identification of dexamethasone-responsive genes by microarray
    Article Snippet: After second-strand synthesis in the presence of DNA polymerase, Escherichia coli RNase H, and E coli DNA ligase (Invitrogen, Cleveland, OH), the DNA was polished with T4 DNA polymerase and purified by phenol/chloroform extraction followed by chromatography on a BioGel P-6 MicroSpin Column (BioRad, Hercules, CA). .. After RNAse H addition and annealing of oligo(dT)-T7 primer, second strand was synthesized without E coli ligase.

    Construct:

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Step 4: Verify correct assembly of the planned yTREX construct in silico using appropriate software (e.g. Clone Manager from Sci‐Ed Software), checking integrity of coding sequences, presence of RBSs and completeness of genes (start to stop codon). .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    SYBR Green Assay:

    Article Title: BH3-only protein Bim more critical than Puma in tyrosine kinase inhibitor-induced apoptosis of human leukemic cells and transduced hematopoietic progenitors carrying oncogenic FLT3
    Article Snippet: .. Quantitative PCR was performed in 10 μL with 2× SYBR Green Mastermix (Applied Biosystems, Warrington, United Kingdom) containing DNA polymerase, dNTPs, buffer, 1 μM each forward and reverse primers, and 4-12 ng template using the 7500 FAST Real-Time System (Applied Biosystems). ..

    Microarray:

    Article Title: Analysis of daily and circadian gene expression in the rat pineal gland
    Article Snippet: In brief, 10 μg each RNA sample was used to synthesize the microarray RNA targets. .. RNA was reverse-transcribed into double-stranded cDNA with a T7 promoter-containing primer using Superscript II, RNase H, and DNA polymerase (Invitrogen).

    Incubation:

    Article Title: Correlation of intestinal disaccharidase activities with the C/T-13910 variant and age
    Article Snippet: Typically, 50 μL of the minisequencing reaction mixture contained 10 pmoles of the minisequencing primers for C/T-13910 and 0.1 μL of either 3H-dCTP corresponding to the lactase non-persistence allele (Amersham, UK) or 3H-dTTP corresponding to the lactase persistence allele, and 0.05 units of DNA polymerase (Dynazyme II, Finnzymes). .. The microtitre plates were incubated for 20 min at 50°C, and the wells were washed.

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: .. Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr. .. The samples were blunt ended by addition of 2 μl of T4 DNA polymerase (Invitrogen) and incubation at 16° for 15 min. Ethanol precipitation was performed (70 μl 5 m NH4 OAc, 650 μl chilled 100% ethanol, 2 μl glycogen) and the sample was resuspended in 8 μl of RNase-free water.

    Article Title: The C/C−13910 genotype of adult-type hypolactasia is associated with an increased risk of colorectal cancer in the Finnish population
    Article Snippet: The minisequencing reaction contained 10 pmol of the minisequencing primer for C/T−13910 (5′-GGC AAT ACA GAT AAG ATA ATG TAG-3′) and 0.1 μl of either H-dCTP or H-dTTP (Amersham Biosciences, Little Chalfont, Buckinghamshire, UK), and 0.05 U of DNA polymerase (Dynazyme II, Finnzymes, Espoo, Finland). .. The microtitre wells were incubated for 15 minutes at 56°C and washed.

    Article Title: Dedicator of cytokinesis 8 is disrupted in two patients with mental retardation and developmental disabilities
    Article Snippet: DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. Slides were incubated in 2× SSC at 37°C for 30 minutes, serially dehydrated in 70%, 80%, and 95% ethanol at room temperature, denatured in 70% formamide/2 × SSC at 72°C for two minutes, then serially dehydrated in 70%, 80%, 95% ethanol.

    In Silico:

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Step 4: Verify correct assembly of the planned yTREX construct in silico using appropriate software (e.g. Clone Manager from Sci‐Ed Software), checking integrity of coding sequences, presence of RBSs and completeness of genes (start to stop codon). .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Expressing:

    Article Title: BH3-only protein Bim more critical than Puma in tyrosine kinase inhibitor-induced apoptosis of human leukemic cells and transduced hematopoietic progenitors carrying oncogenic FLT3
    Article Snippet: Quantitative PCR was performed in 10 μL with 2× SYBR Green Mastermix (Applied Biosystems, Warrington, United Kingdom) containing DNA polymerase, dNTPs, buffer, 1 μM each forward and reverse primers, and 4-12 ng template using the 7500 FAST Real-Time System (Applied Biosystems). .. Relative expression was calculated using the 2−ΔΔCt method.

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: The embryonic stem (ES) cell data sets were downloaded from the Gene Expression Omnibus (GEO) repository (unamplified samples 1–8; ) ( ). .. Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr.

    Article Title: Overexpression of miR-361-5p in triple-negative breast cancer (TNBC) inhibits migration and invasion by targeting RQCD1 and inhibiting the EGFR/PI3K/Akt pathway
    Article Snippet: Pyrobest DNA Polymerase and M-MLV Reverse Transcriptase (Thermo Fisher Scientific, USA) were used for complementary DNA (cDNA) synthesis and PCR reaction. .. PCR conditions for cDNA synthesis were set at 95°C for 15 minutes, and for PCR reaction at 96°C for 5 minutes (initiation), 94°C for 30 seconds (denaturation), 65°C for 30 seconds (annealing), and 75°C for 1 minute (extension) for 33 cycles. miR-361-5p expression was normalized to U6 small nuclear RNA and mRNA expression of selected proteins to the GAPDH gene.

    Modification:

    Article Title: An RNA interference model of RPS19 deficiency in Diamond-Blackfan anemia recapitulates defective hematopoiesis and rescue by dexamethasone: identification of dexamethasone-responsive genes by microarray
    Article Snippet: RNA was linearly amplified by 2 rounds of in vitro transcription (IVT) by the method of Hunter and colleagues, modified to maintain representation of mRNA levels from small starting quantities of RNA and avoid template-independent amplification (Baugh et al ). .. After second-strand synthesis in the presence of DNA polymerase, Escherichia coli RNase H, and E coli DNA ligase (Invitrogen, Cleveland, OH), the DNA was polished with T4 DNA polymerase and purified by phenol/chloroform extraction followed by chromatography on a BioGel P-6 MicroSpin Column (BioRad, Hercules, CA).

    Hybridization:

    Article Title: Analysis of daily and circadian gene expression in the rat pineal gland
    Article Snippet: RNA was reverse-transcribed into double-stranded cDNA with a T7 promoter-containing primer using Superscript II, RNase H, and DNA polymerase (Invitrogen). .. Resulting target cRNA was collected on RNeasy columns (QIAGEN, Valencia, CA, USA), and then fragmented in alkaline buffer for hybridization to the microarray GeneChips.

    Sequencing:

    Article Title: Single-amplicon MSH2 A636P Mutation Testing in Ashkenazi Jewish Patients With Colorectal Cancer
    Article Snippet: The PCR amplification was performed in a final volume of 50 μL, containing 50 ng of DNA, 2.5 mM dNTPs, 15 pmoles of each primer, and 1.25 U of DNA polymerase (AmpliTaq Gold, Applied Biosystems) in Buffer II. .. The purified PCR products were sequenced using the forward and reverse PCR primers in separate sequencing reactions performed with ABI Big Dye Terminator chemistry.

    Article Title: Dedicator of cytokinesis 8 is disrupted in two patients with mental retardation and developmental disabilities
    Article Snippet: Genomic contigs, clones, markers, mapping and sequence information were obtained from the NCBI ( ) databases. .. DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies).

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: An exemplary assembly scheme of a recombinant yTREX vector carrying the partial phz gene cluster from Pseudomonas aeruginosa and the β ‐galactosidase‐encoding lacZ reporter gene (for DNA sequence see Table ) including details on PCR primer sequences which contain the homologous regions for yeast recombineering is shown in Fig. . .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Article Title: Gene Conversion Between Direct Noncoding Repeats Promotes Genetic and Phenotypic Diversity at a Regulatory Locus of Zea mays (L.)
    Article Snippet: Paragraph title: Amplification and sequencing of the p1 noncoding regions: ... The PCR reactions were performed using enhanced DNA polymerase, Elongase (Invitrogen, Carlsbad, CA) with ∼1 min of extension time for every 1-kb fragment size.

    Article Title: Identification of ectodysplasin-A receptor gene deletion at 2q12.2 and a potential autosomal MR locus
    Article Snippet: Genomic contigs, clones, markers, mapping, and sequence information were obtained from the NCBI databases ( ). .. DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies).

    Chromatography:

    Article Title: An RNA interference model of RPS19 deficiency in Diamond-Blackfan anemia recapitulates defective hematopoiesis and rescue by dexamethasone: identification of dexamethasone-responsive genes by microarray
    Article Snippet: .. After second-strand synthesis in the presence of DNA polymerase, Escherichia coli RNase H, and E coli DNA ligase (Invitrogen, Cleveland, OH), the DNA was polished with T4 DNA polymerase and purified by phenol/chloroform extraction followed by chromatography on a BioGel P-6 MicroSpin Column (BioRad, Hercules, CA). .. After precipitation, IVT was performed (Ribomax kit; Promega), and the IVT products were purified using a Qiagen RNeasy Mini Column (Qiagen, Valencia, CA).

    Nick Translation:

    Article Title: Dedicator of cytokinesis 8 is disrupted in two patients with mental retardation and developmental disabilities
    Article Snippet: .. DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. Labeled products were combined with a 50× human Cot-1 DNA (Gibco-BRL) and the chromosome-specific labeled centromeric alphoid probe to a final concentration of 20 ng/μl in Hybrisol VII.

    Article Title: Identification of ectodysplasin-A receptor gene deletion at 2q12.2 and a potential autosomal MR locus
    Article Snippet: .. DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. For in situ hybridization, metaphase chromosome spreads were obtained from lymphoblastoid cells using conventional methods, and fluorescence in situ hybridization (FISH) was performed essentially as described previously., Commercially available chromosome-specific labeled centromeric alphoid probes were used to identify specific chromosomes.

    Infection:

    Article Title: Molecularly defined adult-type hypolactasia among working age people with reference to milk consumption and gastrointestinal symptoms
    Article Snippet: The reaction mixture contained 10 pmol of the detection primer (5’-GGCAATACAGATAAGATAATGTAG-3’), 0.1 μL of either 3 H-dCTP or 3 H-dTTP (Amersham Biosciences, Little Chalfont, Buckinghamshire, UK) and 0.05 U of DNA polymerase (Dynazyme II, Finnzymes, Espoo, Finland). .. Two samples of the 1902 were disqualified after being found infected with hepatitis B.

    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: Expression of Inwardly Rectifying Potassium Channel Subunits 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 Kir subunits (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: Analysis of daily and circadian gene expression in the rat pineal gland
    Article Snippet: Biotin-labeled cRNA probes were generated according to the manufacturer’s protocol (Affymetrix, Santa Clara, CA, USA). .. RNA was reverse-transcribed into double-stranded cDNA with a T7 promoter-containing primer using Superscript II, RNase H, and DNA polymerase (Invitrogen).

    DNA Sequencing:

    Article Title: Gene Conversion Between Direct Noncoding Repeats Promotes Genetic and Phenotypic Diversity at a Regulatory Locus of Zea mays (L.)
    Article Snippet: The PCR reactions were performed using enhanced DNA polymerase, Elongase (Invitrogen, Carlsbad, CA) with ∼1 min of extension time for every 1-kb fragment size. .. Sequencing reactions were provided by the Iowa State University DNA Sequencing Facility.

    Reverse Transcription Polymerase Chain Reaction:

    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.

    Article Title: Expression of Inwardly Rectifying Potassium Channel Subunits 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 Kir subunits (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: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: An exemplary assembly scheme of a recombinant yTREX vector carrying the partial phz gene cluster from Pseudomonas aeruginosa and the β ‐galactosidase‐encoding lacZ reporter gene (for DNA sequence see Table ) including details on PCR primer sequences which contain the homologous regions for yeast recombineering is shown in Fig. . .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Radioactivity:

    Article Title: Correlation of intestinal disaccharidase activities with the C/T-13910 variant and age
    Article Snippet: Typically, 50 μL of the minisequencing reaction mixture contained 10 pmoles of the minisequencing primers for C/T-13910 and 0.1 μL of either 3H-dCTP corresponding to the lactase non-persistence allele (Amersham, UK) or 3H-dTTP corresponding to the lactase persistence allele, and 0.05 units of DNA polymerase (Dynazyme II, Finnzymes). .. The detection primer was eluted, and the eluted radioactivity was measured in a liquid scintillation counter (Rackbeta 1209, Wallac, Finland).

    Article Title: The C/C−13910 genotype of adult-type hypolactasia is associated with an increased risk of colorectal cancer in the Finnish population
    Article Snippet: The minisequencing reaction contained 10 pmol of the minisequencing primer for C/T−13910 (5′-GGC AAT ACA GAT AAG ATA ATG TAG-3′) and 0.1 μl of either H-dCTP or H-dTTP (Amersham Biosciences, Little Chalfont, Buckinghamshire, UK), and 0.05 U of DNA polymerase (Dynazyme II, Finnzymes, Espoo, Finland). .. The detection primer was eluted, and radioactivity measured in a liquid scintillation counter (Rackbeta 1209; Wallac, Turku, Finland).

    Article Title: Molecularly defined adult-type hypolactasia among working age people with reference to milk consumption and gastrointestinal symptoms
    Article Snippet: The reaction mixture contained 10 pmol of the detection primer (5’-GGCAATACAGATAAGATAATGTAG-3’), 0.1 μL of either 3 H-dCTP or 3 H-dTTP (Amersham Biosciences, Little Chalfont, Buckinghamshire, UK) and 0.05 U of DNA polymerase (Dynazyme II, Finnzymes, Espoo, Finland). .. Finally, the attached detection primer was eluted by NaOH treatment and the radioactivity measured in a liquid scintillation counter (Rackbeta 1209; Wallac, Turku, Finland).

    Fluorescence:

    Article Title: Identification of ectodysplasin-A receptor gene deletion at 2q12.2 and a potential autosomal MR locus
    Article Snippet: DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. For in situ hybridization, metaphase chromosome spreads were obtained from lymphoblastoid cells using conventional methods, and fluorescence in situ hybridization (FISH) was performed essentially as described previously., Commercially available chromosome-specific labeled centromeric alphoid probes were used to identify specific chromosomes.

    Mutagenesis:

    Article Title: Single-amplicon MSH2 A636P Mutation Testing in Ashkenazi Jewish Patients With Colorectal Cancer
    Article Snippet: Paragraph title: Mutation Analysis ... The PCR amplification was performed in a final volume of 50 μL, containing 50 ng of DNA, 2.5 mM dNTPs, 15 pmoles of each primer, and 1.25 U of DNA polymerase (AmpliTaq Gold, Applied Biosystems) in Buffer II.

    Isolation:

    Article Title: Correlation of intestinal disaccharidase activities with the C/T-13910 variant and age
    Article Snippet: DNA was isolated from intestinal biopsy specimens by phenol-chloroform extraction according to standard procedures[ ]. .. Typically, 50 μL of the minisequencing reaction mixture contained 10 pmoles of the minisequencing primers for C/T-13910 and 0.1 μL of either 3H-dCTP corresponding to the lactase non-persistence allele (Amersham, UK) or 3H-dTTP corresponding to the lactase persistence allele, and 0.05 units of DNA polymerase (Dynazyme II, Finnzymes).

    Article Title: BH3-only protein Bim more critical than Puma in tyrosine kinase inhibitor-induced apoptosis of human leukemic cells and transduced hematopoietic progenitors carrying oncogenic FLT3
    Article Snippet: Total RNA was isolated using RNeasy mini kit (Qiagen, Hilden, Germany) and reversed transcribed to cDNA by SuperScript III Reverse Transcriptase (Invitrogen). .. Quantitative PCR was performed in 10 μL with 2× SYBR Green Mastermix (Applied Biosystems, Warrington, United Kingdom) containing DNA polymerase, dNTPs, buffer, 1 μM each forward and reverse primers, and 4-12 ng template using the 7500 FAST Real-Time System (Applied Biosystems).

    Article Title: Dedicator of cytokinesis 8 is disrupted in two patients with mental retardation and developmental disabilities
    Article Snippet: .. DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. Labeled products were combined with a 50× human Cot-1 DNA (Gibco-BRL) and the chromosome-specific labeled centromeric alphoid probe to a final concentration of 20 ng/μl in Hybrisol VII.

    Article Title: Identification of ectodysplasin-A receptor gene deletion at 2q12.2 and a potential autosomal MR locus
    Article Snippet: .. DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. For in situ hybridization, metaphase chromosome spreads were obtained from lymphoblastoid cells using conventional methods, and fluorescence in situ hybridization (FISH) was performed essentially as described previously., Commercially available chromosome-specific labeled centromeric alphoid probes were used to identify specific chromosomes.

    Article Title: Analysis of daily and circadian gene expression in the rat pineal gland
    Article Snippet: Total cellular RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s protocol. .. RNA was reverse-transcribed into double-stranded cDNA with a T7 promoter-containing primer using Superscript II, RNase H, and DNA polymerase (Invitrogen).

    Labeling:

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: Adult mouse tissues were dissected from six week FVB mice and labeled per manufacturer's specifications without amplification. .. Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr.

    Article Title: Dedicator of cytokinesis 8 is disrupted in two patients with mental retardation and developmental disabilities
    Article Snippet: .. DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. Labeled products were combined with a 50× human Cot-1 DNA (Gibco-BRL) and the chromosome-specific labeled centromeric alphoid probe to a final concentration of 20 ng/μl in Hybrisol VII.

    Article Title: Identification of ectodysplasin-A receptor gene deletion at 2q12.2 and a potential autosomal MR locus
    Article Snippet: .. DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. For in situ hybridization, metaphase chromosome spreads were obtained from lymphoblastoid cells using conventional methods, and fluorescence in situ hybridization (FISH) was performed essentially as described previously., Commercially available chromosome-specific labeled centromeric alphoid probes were used to identify specific chromosomes.

    Mouse Assay:

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: KitlSl /KitlSl-d male mice were purchased from the Jackson Laboratories. .. Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr.

    Polymerase Chain Reaction:

    Article Title: Correlation of intestinal disaccharidase activities with the C/T-13910 variant and age
    Article Snippet: A 10-μL aliquot of the PCR product was captured in a streptavidin-coated microtitre well (Labsystems, Finland). .. Typically, 50 μL of the minisequencing reaction mixture contained 10 pmoles of the minisequencing primers for C/T-13910 and 0.1 μL of either 3H-dCTP corresponding to the lactase non-persistence allele (Amersham, UK) or 3H-dTTP corresponding to the lactase persistence allele, and 0.05 units of DNA polymerase (Dynazyme II, Finnzymes).

    Article Title: Single-amplicon MSH2 A636P Mutation Testing in Ashkenazi Jewish Patients With Colorectal Cancer
    Article Snippet: .. The PCR amplification was performed in a final volume of 50 μL, containing 50 ng of DNA, 2.5 mM dNTPs, 15 pmoles of each primer, and 1.25 U of DNA polymerase (AmpliTaq Gold, Applied Biosystems) in Buffer II. .. The PCR amplification consisted of 10 minutes at 95°C, followed by 35 cycles of 30 seconds at 94°C, 30 seconds at 55°C, and 45 seconds at 72°C; a final extension of 7 minutes ended the amplification reaction.

    Article Title: The C/C−13910 genotype of adult-type hypolactasia is associated with an increased risk of colorectal cancer in the Finnish population
    Article Snippet: The PCR product (10 μl) was captured in a streptavidin coated microtitre well (Thermo Electron, Helsinki, Finland); two parallel minisequencing reactions were carried out for each PCR product. .. The minisequencing reaction contained 10 pmol of the minisequencing primer for C/T−13910 (5′-GGC AAT ACA GAT AAG ATA ATG TAG-3′) and 0.1 μl of either H-dCTP or H-dTTP (Amersham Biosciences, Little Chalfont, Buckinghamshire, UK), and 0.05 U of DNA polymerase (Dynazyme II, Finnzymes, Espoo, Finland).

    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: Molecularly defined adult-type hypolactasia among working age people with reference to milk consumption and gastrointestinal symptoms
    Article Snippet: Briefly, in the mini-sequencing of the C/T-13910 single nucleotide polymorphism, two 10-μL aliquots of the biotin-labeled PCR product were captured to streptavidin coated microtitre wells (Thermo Electron, Helsinki, Finland). .. The reaction mixture contained 10 pmol of the detection primer (5’-GGCAATACAGATAAGATAATGTAG-3’), 0.1 μL of either 3 H-dCTP or 3 H-dTTP (Amersham Biosciences, Little Chalfont, Buckinghamshire, UK) and 0.05 U of DNA polymerase (Dynazyme II, Finnzymes, Espoo, Finland).

    Article Title: Expression of Inwardly Rectifying Potassium Channel Subunits 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 Kir subunits (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: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH). .. Use a PCR programme according to the manufacturer's manual.

    Article Title: Gene Conversion Between Direct Noncoding Repeats Promotes Genetic and Phenotypic Diversity at a Regulatory Locus of Zea mays (L.)
    Article Snippet: .. The PCR reactions were performed using enhanced DNA polymerase, Elongase (Invitrogen, Carlsbad, CA) with ∼1 min of extension time for every 1-kb fragment size. .. Optimal PCR parameters were followed as suggested by the manufacturer.

    Article Title: Overexpression of miR-361-5p in triple-negative breast cancer (TNBC) inhibits migration and invasion by targeting RQCD1 and inhibiting the EGFR/PI3K/Akt pathway
    Article Snippet: .. Pyrobest DNA Polymerase and M-MLV Reverse Transcriptase (Thermo Fisher Scientific, USA) were used for complementary DNA (cDNA) synthesis and PCR reaction. .. PCR conditions for cDNA synthesis were set at 95°C for 15 minutes, and for PCR reaction at 96°C for 5 minutes (initiation), 94°C for 30 seconds (denaturation), 65°C for 30 seconds (annealing), and 75°C for 1 minute (extension) for 33 cycles. miR-361-5p expression was normalized to U6 small nuclear RNA and mRNA expression of selected proteins to the GAPDH gene.

    Quantitative RT-PCR:

    Article Title: Overexpression of miR-361-5p in triple-negative breast cancer (TNBC) inhibits migration and invasion by targeting RQCD1 and inhibiting the EGFR/PI3K/Akt pathway
    Article Snippet: Paragraph title: qRT-PCR ... Pyrobest DNA Polymerase and M-MLV Reverse Transcriptase (Thermo Fisher Scientific, USA) were used for complementary DNA (cDNA) synthesis and PCR reaction.

    Activated Clotting Time Assay:

    Article Title: The C/C−13910 genotype of adult-type hypolactasia is associated with an increased risk of colorectal cancer in the Finnish population
    Article Snippet: The DNA fragment spanning the C/T−13910 variant was amplified using one biotinylated (5′-CCT CGT TAA TAC CCA CTG ACC TA-3′) primer and one unbiotinylated (5′-GTC ACT TTG ATA TGA TGA GAG CA-3′) primer. .. The minisequencing reaction contained 10 pmol of the minisequencing primer for C/T−13910 (5′-GGC AAT ACA GAT AAG ATA ATG TAG-3′) and 0.1 μl of either H-dCTP or H-dTTP (Amersham Biosciences, Little Chalfont, Buckinghamshire, UK), and 0.05 U of DNA polymerase (Dynazyme II, Finnzymes, Espoo, Finland).

    Purification:

    Article Title: Single-amplicon MSH2 A636P Mutation Testing in Ashkenazi Jewish Patients With Colorectal Cancer
    Article Snippet: The PCR amplification was performed in a final volume of 50 μL, containing 50 ng of DNA, 2.5 mM dNTPs, 15 pmoles of each primer, and 1.25 U of DNA polymerase (AmpliTaq Gold, Applied Biosystems) in Buffer II. .. PCR products were purified with the QIAquick PCR purification kit (Qiagen) and quantified by agarose gel electrophoresis.

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr. .. Samples were purified using RNeasy mini kit (QIAGEN), eluted in 50 μl H2 O, quantitated by OD260 , and 400 ng cRNA in 12 μl H2 O was subjected to a second round of amplification.

    Article Title: Gene Conversion Between Direct Noncoding Repeats Promotes Genetic and Phenotypic Diversity at a Regulatory Locus of Zea mays (L.)
    Article Snippet: The PCR reactions were performed using enhanced DNA polymerase, Elongase (Invitrogen, Carlsbad, CA) with ∼1 min of extension time for every 1-kb fragment size. .. To minimize the problems caused by PCR artifacts due to annealing of partial extension products to homologous regions in the template DNA pool, the PCR products from three independent reactions were mixed, purified using a gel extraction kit (QIAGEN, Valencia, CA), and sequenced directly from both directions.

    Article Title: An RNA interference model of RPS19 deficiency in Diamond-Blackfan anemia recapitulates defective hematopoiesis and rescue by dexamethasone: identification of dexamethasone-responsive genes by microarray
    Article Snippet: .. After second-strand synthesis in the presence of DNA polymerase, Escherichia coli RNase H, and E coli DNA ligase (Invitrogen, Cleveland, OH), the DNA was polished with T4 DNA polymerase and purified by phenol/chloroform extraction followed by chromatography on a BioGel P-6 MicroSpin Column (BioRad, Hercules, CA). .. After precipitation, IVT was performed (Ribomax kit; Promega), and the IVT products were purified using a Qiagen RNeasy Mini Column (Qiagen, Valencia, CA).

    In Situ Hybridization:

    Article Title: Dedicator of cytokinesis 8 is disrupted in two patients with mental retardation and developmental disabilities
    Article Snippet: DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. For in situ hybridization, the metaphase chromosome spreads were obtained from lympoblastoid cells (CMS6482) or from fibroblast cells (CMS1485) using conventional methods.

    Article Title: Identification of ectodysplasin-A receptor gene deletion at 2q12.2 and a potential autosomal MR locus
    Article Snippet: DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. For in situ hybridization, metaphase chromosome spreads were obtained from lymphoblastoid cells using conventional methods, and fluorescence in situ hybridization (FISH) was performed essentially as described previously., Commercially available chromosome-specific labeled centromeric alphoid probes were used to identify specific chromosomes.

    Plasmid Preparation:

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Paragraph title: Generation of gene cluster DNA fragments and yeast assembly cloning in the yTREX vector ... Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Software:

    Article Title: BH3-only protein Bim more critical than Puma in tyrosine kinase inhibitor-induced apoptosis of human leukemic cells and transduced hematopoietic progenitors carrying oncogenic FLT3
    Article Snippet: Quantitative PCR was performed in 10 μL with 2× SYBR Green Mastermix (Applied Biosystems, Warrington, United Kingdom) containing DNA polymerase, dNTPs, buffer, 1 μM each forward and reverse primers, and 4-12 ng template using the 7500 FAST Real-Time System (Applied Biosystems). .. Primer sequences designed with Primer express software (Applied Biosystems) were as follows: GusB forward primer: 5′-TGGTTGGAGAGCT-CATTTGGA-3′ and reverse primer: 5′-ACTCTCGTCGGTGACTGT-TCAG-3′; Bim forward primer: 5′-TGGCAAAGCAACCTTCTGATG-3′ and reverse primer: 5′-GCAGGCTGCAATTGTCTACCT-3′; and Puma forward primer: 5′-GAAGAGCAAATGAGCCAAACG-3′ and reverse primer: 5′-GGAGCAACCGGCAAACG-3′.

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Step 4: Verify correct assembly of the planned yTREX construct in silico using appropriate software (e.g. Clone Manager from Sci‐Ed Software), checking integrity of coding sequences, presence of RBSs and completeness of genes (start to stop codon). .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Real-time Polymerase Chain Reaction:

    Article Title: BH3-only protein Bim more critical than Puma in tyrosine kinase inhibitor-induced apoptosis of human leukemic cells and transduced hematopoietic progenitors carrying oncogenic FLT3
    Article Snippet: .. Quantitative PCR was performed in 10 μL with 2× SYBR Green Mastermix (Applied Biosystems, Warrington, United Kingdom) containing DNA polymerase, dNTPs, buffer, 1 μM each forward and reverse primers, and 4-12 ng template using the 7500 FAST Real-Time System (Applied Biosystems). ..

    Binding Assay:

    Article Title: An RNA interference model of RPS19 deficiency in Diamond-Blackfan anemia recapitulates defective hematopoiesis and rescue by dexamethasone: identification of dexamethasone-responsive genes by microarray
    Article Snippet: Briefly, first-round amplification was carried out with oligo(dT)-T7 primers and Superscript II (Invitrogen) in the presence of the single-stranded nucleic acid binding protein T4gp32 (USB) for one hour at 4°C. .. After second-strand synthesis in the presence of DNA polymerase, Escherichia coli RNase H, and E coli DNA ligase (Invitrogen, Cleveland, OH), the DNA was polished with T4 DNA polymerase and purified by phenol/chloroform extraction followed by chromatography on a BioGel P-6 MicroSpin Column (BioRad, Hercules, CA).

    Agarose Gel Electrophoresis:

    Article Title: Single-amplicon MSH2 A636P Mutation Testing in Ashkenazi Jewish Patients With Colorectal Cancer
    Article Snippet: The PCR amplification was performed in a final volume of 50 μL, containing 50 ng of DNA, 2.5 mM dNTPs, 15 pmoles of each primer, and 1.25 U of DNA polymerase (AmpliTaq Gold, Applied Biosystems) in Buffer II. .. PCR products were purified with the QIAquick PCR purification kit (Qiagen) and quantified by agarose gel electrophoresis.

    In Vitro:

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr. .. In vitro transcription was performed with the MEGAscript high-yield transcription kit (Ambion) and incubated at 37° for 4 hr.

    Article Title: An RNA interference model of RPS19 deficiency in Diamond-Blackfan anemia recapitulates defective hematopoiesis and rescue by dexamethasone: identification of dexamethasone-responsive genes by microarray
    Article Snippet: RNA was linearly amplified by 2 rounds of in vitro transcription (IVT) by the method of Hunter and colleagues, modified to maintain representation of mRNA levels from small starting quantities of RNA and avoid template-independent amplification (Baugh et al ). .. After second-strand synthesis in the presence of DNA polymerase, Escherichia coli RNase H, and E coli DNA ligase (Invitrogen, Cleveland, OH), the DNA was polished with T4 DNA polymerase and purified by phenol/chloroform extraction followed by chromatography on a BioGel P-6 MicroSpin Column (BioRad, Hercules, CA).

    Ethanol Precipitation:

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr. .. The samples were blunt ended by addition of 2 μl of T4 DNA polymerase (Invitrogen) and incubation at 16° for 15 min. Ethanol precipitation was performed (70 μl 5 m NH4 OAc, 650 μl chilled 100% ethanol, 2 μl glycogen) and the sample was resuspended in 8 μl of RNase-free water.

    Article Title: Analysis of daily and circadian gene expression in the rat pineal gland
    Article Snippet: RNA was reverse-transcribed into double-stranded cDNA with a T7 promoter-containing primer using Superscript II, RNase H, and DNA polymerase (Invitrogen). .. After extraction with phenol-chloroform and ethanol precipitation with ammonium acetate, the cDNA was used as a template to generate biotin-labeled cRNA probe (Enzo Bioarray, Affymetrix, Santa Clara, CA, USA).

    Random Hexamer Labeling:

    Article Title: An RNA interference model of RPS19 deficiency in Diamond-Blackfan anemia recapitulates defective hematopoiesis and rescue by dexamethasone: identification of dexamethasone-responsive genes by microarray
    Article Snippet: After second-strand synthesis in the presence of DNA polymerase, Escherichia coli RNase H, and E coli DNA ligase (Invitrogen, Cleveland, OH), the DNA was polished with T4 DNA polymerase and purified by phenol/chloroform extraction followed by chromatography on a BioGel P-6 MicroSpin Column (BioRad, Hercules, CA). .. For second-round amplification, random hexamer primers were added to the cRNA and RT was carried out at 37°C (20 minutes), 42°C (20 minutes), 50°C (10 minutes), and 55°C (10 minutes), and heat inactivated at 65°C (15 minutes).

    Spectrophotometry:

    Article Title: Overexpression of miR-361-5p in triple-negative breast cancer (TNBC) inhibits migration and invasion by targeting RQCD1 and inhibiting the EGFR/PI3K/Akt pathway
    Article Snippet: The concentration of RNA was measured with a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, USA). .. Pyrobest DNA Polymerase and M-MLV Reverse Transcriptase (Thermo Fisher Scientific, USA) were used for complementary DNA (cDNA) synthesis and PCR reaction.

    Concentration Assay:

    Article Title: Dedicator of cytokinesis 8 is disrupted in two patients with mental retardation and developmental disabilities
    Article Snippet: DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. Labeled products were combined with a 50× human Cot-1 DNA (Gibco-BRL) and the chromosome-specific labeled centromeric alphoid probe to a final concentration of 20 ng/μl in Hybrisol VII.

    Article Title: Overexpression of miR-361-5p in triple-negative breast cancer (TNBC) inhibits migration and invasion by targeting RQCD1 and inhibiting the EGFR/PI3K/Akt pathway
    Article Snippet: The concentration of RNA was measured with a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, USA). .. Pyrobest DNA Polymerase and M-MLV Reverse Transcriptase (Thermo Fisher Scientific, USA) were used for complementary DNA (cDNA) synthesis and PCR reaction.

    CTG Assay:

    Article Title: The C/C−13910 genotype of adult-type hypolactasia is associated with an increased risk of colorectal cancer in the Finnish population
    Article Snippet: The DNA fragment spanning the C/T−13910 variant was amplified using one biotinylated (5′-CCT CGT TAA TAC CCA CTG ACC TA-3′) primer and one unbiotinylated (5′-GTC ACT TTG ATA TGA TGA GAG CA-3′) primer. .. The minisequencing reaction contained 10 pmol of the minisequencing primer for C/T−13910 (5′-GGC AAT ACA GAT AAG ATA ATG TAG-3′) and 0.1 μl of either H-dCTP or H-dTTP (Amersham Biosciences, Little Chalfont, Buckinghamshire, UK), and 0.05 U of DNA polymerase (Dynazyme II, Finnzymes, Espoo, Finland).

    Gel Extraction:

    Article Title: Gene Conversion Between Direct Noncoding Repeats Promotes Genetic and Phenotypic Diversity at a Regulatory Locus of Zea mays (L.)
    Article Snippet: The PCR reactions were performed using enhanced DNA polymerase, Elongase (Invitrogen, Carlsbad, CA) with ∼1 min of extension time for every 1-kb fragment size. .. To minimize the problems caused by PCR artifacts due to annealing of partial extension products to homologous regions in the template DNA pool, the PCR products from three independent reactions were mixed, purified using a gel extraction kit (QIAGEN, Valencia, CA), and sequenced directly from both directions.

    Variant Assay:

    Article Title: Correlation of intestinal disaccharidase activities with the C/T-13910 variant and age
    Article Snippet: The DNA fragment spanning the C/T-13910 variant was amplified using a biotinylated and an unbiotinylated primer (primers available upon request). .. Typically, 50 μL of the minisequencing reaction mixture contained 10 pmoles of the minisequencing primers for C/T-13910 and 0.1 μL of either 3H-dCTP corresponding to the lactase non-persistence allele (Amersham, UK) or 3H-dTTP corresponding to the lactase persistence allele, and 0.05 units of DNA polymerase (Dynazyme II, Finnzymes).

    Article Title: The C/C−13910 genotype of adult-type hypolactasia is associated with an increased risk of colorectal cancer in the Finnish population
    Article Snippet: The DNA fragment spanning the C/T−13910 variant was amplified using one biotinylated (5′-CCT CGT TAA TAC CCA CTG ACC TA-3′) primer and one unbiotinylated (5′-GTC ACT TTG ATA TGA TGA GAG CA-3′) primer. .. The minisequencing reaction contained 10 pmol of the minisequencing primer for C/T−13910 (5′-GGC AAT ACA GAT AAG ATA ATG TAG-3′) and 0.1 μl of either H-dCTP or H-dTTP (Amersham Biosciences, Little Chalfont, Buckinghamshire, UK), and 0.05 U of DNA polymerase (Dynazyme II, Finnzymes, Espoo, Finland).

    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.

    Fluorescence In Situ Hybridization:

    Article Title: Identification of ectodysplasin-A receptor gene deletion at 2q12.2 and a potential autosomal MR locus
    Article Snippet: DNA was isolated using Qiagen Mini-Prep columns and was labeled by incorporation of digoxigenin-11-dUTP or biotin-16-dUTP (Boehringer Mannheim) by nick translation using DNA polymerase (Life Technologies). .. For in situ hybridization, metaphase chromosome spreads were obtained from lymphoblastoid cells using conventional methods, and fluorescence in situ hybridization (FISH) was performed essentially as described previously., Commercially available chromosome-specific labeled centromeric alphoid probes were used to identify specific chromosomes.

    Homologous Recombination:

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Such homologies could result in false pairing during homologous recombination. .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

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  • 90
    Thermo Fisher platinum superfi dna polymerase
    Clustering of samples amplified with two different polymerases on a non-metric multidimensional scaling (NMDS) plot of the Bray-Curtis dissimilarities. Points are colored by applied extraction kit. The encircled pairs correspond to a single sample where each data point represents one 16 S rRNA amplification with Phusion Hot Start II High-Fidelity <t>DNA</t> Polymerase and the another with Platinum <t>SuperFi</t> DNA Polymerase. Sample pairs labeled with * were stored in PBS.
    Platinum Superfi Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/platinum superfi dna polymerase/product/Thermo Fisher
    Average 90 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    platinum superfi dna polymerase - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    99
    Thermo Fisher dna polymerases
    Genotyping of Arabidopsis thaliana plants from SALK_100012 line with (a) <t>Phusion</t> High-Fidelity <t>DNA</t> polymerase; (b) Ex-Taq DNA polymerase; (c) Taq DNA polymerase; (d) Emerald Amp GT PCR Master Mix. WT - DNA from wild-type Arabidopsis plant (positive control for wild-type PCR band); 1–8 - individual plants from SALK_100012 line. Molecular weight DNA standards are shown on each side. Loading order for each DNA sample: PCR reaction for wild-type allele, PCR reaction for T-DNA mutant allele.
    Dna Polymerases, 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/dna polymerases/product/Thermo Fisher
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    dna polymerases - by Bioz Stars, 2020-02
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    90
    Thermo Fisher phi29 dna polymerase
    Size distribution of <t>DNA-magnesium-pyrophosphate</t> particles as a function of multiple displacement amplification reaction times, with and without the heat-inactivation step. At different time points (17, 38 and 66 h) of the MDA reaction in 3-pL droplets at 30 °C, the resulting DNA-Mg-PP i particles were released from droplets and imaged under TEM to measure their size. The first set of measurements (green) was made without heat-inactivation step. The second set of measurements (grey) was made with heat-inactivation step at 65 °C for 15 min that is typically used to inactivate <t>phi29</t> DNA polymerase and terminate the MDA reaction.
    Phi29 Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phi29 dna polymerase/product/Thermo Fisher
    Average 90 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    phi29 dna polymerase - by Bioz Stars, 2020-02
    90/100 stars
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    Image Search Results


    Clustering of samples amplified with two different polymerases on a non-metric multidimensional scaling (NMDS) plot of the Bray-Curtis dissimilarities. Points are colored by applied extraction kit. The encircled pairs correspond to a single sample where each data point represents one 16 S rRNA amplification with Phusion Hot Start II High-Fidelity DNA Polymerase and the another with Platinum SuperFi DNA Polymerase. Sample pairs labeled with * were stored in PBS.

    Journal: Scientific Reports

    Article Title: The impact of storage buffer, DNA extraction method, and polymerase on microbial analysis

    doi: 10.1038/s41598-018-24573-y

    Figure Lengend Snippet: Clustering of samples amplified with two different polymerases on a non-metric multidimensional scaling (NMDS) plot of the Bray-Curtis dissimilarities. Points are colored by applied extraction kit. The encircled pairs correspond to a single sample where each data point represents one 16 S rRNA amplification with Phusion Hot Start II High-Fidelity DNA Polymerase and the another with Platinum SuperFi DNA Polymerase. Sample pairs labeled with * were stored in PBS.

    Article Snippet: Platinum SuperFi DNA Polymerase (Thermo Fisher Scientific) and the Phusion Hot Start II High-Fidelity DNA Polymerase (Thermo Fisher Scientific) were both tested for amplification.

    Techniques: Amplification, Labeling

    Genotyping of Arabidopsis thaliana plants from SALK_100012 line with (a) Phusion High-Fidelity DNA polymerase; (b) Ex-Taq DNA polymerase; (c) Taq DNA polymerase; (d) Emerald Amp GT PCR Master Mix. WT - DNA from wild-type Arabidopsis plant (positive control for wild-type PCR band); 1–8 - individual plants from SALK_100012 line. Molecular weight DNA standards are shown on each side. Loading order for each DNA sample: PCR reaction for wild-type allele, PCR reaction for T-DNA mutant allele.

    Journal: BioNanoScience

    Article Title: Selection of efficient Taq DNA polymerase to optimize T-DNA genotyping method for rapid detection of mutant Arabidopsis thaliana plants

    doi: 10.1007/s12668-016-0253-6

    Figure Lengend Snippet: Genotyping of Arabidopsis thaliana plants from SALK_100012 line with (a) Phusion High-Fidelity DNA polymerase; (b) Ex-Taq DNA polymerase; (c) Taq DNA polymerase; (d) Emerald Amp GT PCR Master Mix. WT - DNA from wild-type Arabidopsis plant (positive control for wild-type PCR band); 1–8 - individual plants from SALK_100012 line. Molecular weight DNA standards are shown on each side. Loading order for each DNA sample: PCR reaction for wild-type allele, PCR reaction for T-DNA mutant allele.

    Article Snippet: The following DNA polymerases were tested: Phusion High-Fidelity DNA polymerase (ThermoFisher Scientific, USA, cat #F530S), Ex-Taq DNA polymerase (Clontech, USA, cat #RR001A), Taq DNA polymerase (SibEnzyme, Russia, cat #E331), Emerald Amp GT PCR Master Mix (Clontech, USA, cat #RR310A).

    Techniques: Polymerase Chain Reaction, Positive Control, Molecular Weight, Mutagenesis

    Primase vs. primer-dependent DNA polymerase activity of intact PolpTN2, PolPTNΔ 311–923 and Taq polymerase. ( A ) Intact PolpTN2 (40 ng/µl), ( B ) PolpTN2Δ 311–923 (80 ng/µl) or ( C ) Taq DNA polymerase (0,05 u/µl) were incubated at 70°C in Taq buffer + 0,4 mM dNTP and 2 ng/µl M13mp18 DNA, which was or was not hybridized to the complementary oligonucleotide primer M13 forward ( Supplementary Table S1 ). At t = 0, 5, 10 and 15 min, aliquots were withdrawn and quenched into 25 mM EDTA. The amount of ds DNA synthesized was then determined using Sybr® Green I fluorescence as described in Materials and Methods. Circles, continuous line: synthesis with hybridized primer; squares, dashed line: synthesis without primer. Points are the average of two determinations. The standard deviation is indicated by error bars.

    Journal: Nucleic Acids Research

    Article Title: A highly divergent archaeo-eukaryotic primase from the Thermococcus nautilus plasmid, pTN2

    doi: 10.1093/nar/gkt1385

    Figure Lengend Snippet: Primase vs. primer-dependent DNA polymerase activity of intact PolpTN2, PolPTNΔ 311–923 and Taq polymerase. ( A ) Intact PolpTN2 (40 ng/µl), ( B ) PolpTN2Δ 311–923 (80 ng/µl) or ( C ) Taq DNA polymerase (0,05 u/µl) were incubated at 70°C in Taq buffer + 0,4 mM dNTP and 2 ng/µl M13mp18 DNA, which was or was not hybridized to the complementary oligonucleotide primer M13 forward ( Supplementary Table S1 ). At t = 0, 5, 10 and 15 min, aliquots were withdrawn and quenched into 25 mM EDTA. The amount of ds DNA synthesized was then determined using Sybr® Green I fluorescence as described in Materials and Methods. Circles, continuous line: synthesis with hybridized primer; squares, dashed line: synthesis without primer. Points are the average of two determinations. The standard deviation is indicated by error bars.

    Article Snippet: Non-radioactive primase and DNA polymerase assays Reactions were carried out in Taq polymerase buffer (75 mM Tris-HCl pH 8.8 at 25°C, 20 mM (NH4 )2 SO4 , 0.01% Tween 20) (Thermo) containing 2.5 mM MgCl2 , 0.4 mM of each of the four dNTPs, 2 ng/µl of M13mp18 single-stranded DNA (New England Biolabs) and enzyme components as described in the text.

    Techniques: Activity Assay, Incubation, Synthesized, SYBR Green Assay, Fluorescence, Standard Deviation

    Efficiency of intact PolpTN2 and PolpTN2Δ 311–923 in priming DNA synthesis by T. nautilus PolB and Taq DNA polymerase. T. nautilus PolB (8 ng/µl) or Taq polymerase (0,05 u/µl) were incubated at 70°C in Taq buffer + 0,4 mM dNTP and 2 ng/µl M13mp18 DNA (without annealed primer) with increasing concentrations of either intact PolpTN2 ( A ) or PolpTN2 Δ 311–923 ( B ). DNA synthesis by intact PolpTN2 or PolpTN2 Δ 311–923 alone was included as a control. At t = 0 and 6 min, aliquots were withdrawn, quenched in 25 mM EDTA and assayed for ds DNA using Sybr® Green I fluorescence. Circles, continuous line: PolpTN2 or PolpTN2 Δ 311–923 alone; squares, dashed line: PolpTN2 or PolpTN2 Δ 311–923 plus Taq DNA polymerase; triangles, dotted line: PolpTN2 or PolpTN2 Δ 311–923 plus T. nautilus PolB DNA polymerase. Points are the average of three determinations. The standard deviation is indicated by error bars.

    Journal: Nucleic Acids Research

    Article Title: A highly divergent archaeo-eukaryotic primase from the Thermococcus nautilus plasmid, pTN2

    doi: 10.1093/nar/gkt1385

    Figure Lengend Snippet: Efficiency of intact PolpTN2 and PolpTN2Δ 311–923 in priming DNA synthesis by T. nautilus PolB and Taq DNA polymerase. T. nautilus PolB (8 ng/µl) or Taq polymerase (0,05 u/µl) were incubated at 70°C in Taq buffer + 0,4 mM dNTP and 2 ng/µl M13mp18 DNA (without annealed primer) with increasing concentrations of either intact PolpTN2 ( A ) or PolpTN2 Δ 311–923 ( B ). DNA synthesis by intact PolpTN2 or PolpTN2 Δ 311–923 alone was included as a control. At t = 0 and 6 min, aliquots were withdrawn, quenched in 25 mM EDTA and assayed for ds DNA using Sybr® Green I fluorescence. Circles, continuous line: PolpTN2 or PolpTN2 Δ 311–923 alone; squares, dashed line: PolpTN2 or PolpTN2 Δ 311–923 plus Taq DNA polymerase; triangles, dotted line: PolpTN2 or PolpTN2 Δ 311–923 plus T. nautilus PolB DNA polymerase. Points are the average of three determinations. The standard deviation is indicated by error bars.

    Article Snippet: Non-radioactive primase and DNA polymerase assays Reactions were carried out in Taq polymerase buffer (75 mM Tris-HCl pH 8.8 at 25°C, 20 mM (NH4 )2 SO4 , 0.01% Tween 20) (Thermo) containing 2.5 mM MgCl2 , 0.4 mM of each of the four dNTPs, 2 ng/µl of M13mp18 single-stranded DNA (New England Biolabs) and enzyme components as described in the text.

    Techniques: DNA Synthesis, Incubation, SYBR Green Assay, Fluorescence, Standard Deviation

    Size distribution of DNA-magnesium-pyrophosphate particles as a function of multiple displacement amplification reaction times, with and without the heat-inactivation step. At different time points (17, 38 and 66 h) of the MDA reaction in 3-pL droplets at 30 °C, the resulting DNA-Mg-PP i particles were released from droplets and imaged under TEM to measure their size. The first set of measurements (green) was made without heat-inactivation step. The second set of measurements (grey) was made with heat-inactivation step at 65 °C for 15 min that is typically used to inactivate phi29 DNA polymerase and terminate the MDA reaction.

    Journal: Micromachines

    Article Title: Droplet Microfluidics Approach for Single-DNA Molecule Amplification and Condensation into DNA-Magnesium-Pyrophosphate Particles

    doi: 10.3390/mi8020062

    Figure Lengend Snippet: Size distribution of DNA-magnesium-pyrophosphate particles as a function of multiple displacement amplification reaction times, with and without the heat-inactivation step. At different time points (17, 38 and 66 h) of the MDA reaction in 3-pL droplets at 30 °C, the resulting DNA-Mg-PP i particles were released from droplets and imaged under TEM to measure their size. The first set of measurements (green) was made without heat-inactivation step. The second set of measurements (grey) was made with heat-inactivation step at 65 °C for 15 min that is typically used to inactivate phi29 DNA polymerase and terminate the MDA reaction.

    Article Snippet: In this context, DNA amplification driven by phi29 DNA polymerase provides an alternative approach to amplify long DNA molecules and because of isothermal reaction conditions [ , ], the potential problems associated with emulsion stability become irrelevant.

    Techniques: Multiple Displacement Amplification, Transmission Electron Microscopy