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Bio-Rad dna polymerase
Dna Polymerase, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dna polymerase - by Bioz Stars, 2020-04
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Amplification:

Article Title: Impaired glucose transporter-1 degradation and increased glucose transport and oxidative stress in response to high glucose in chondrocytes from osteoarthritic versus normal human cartilage
Article Snippet: Quantitative real-time RT-PCR was performed with iTaq™ DNA polymerase using iQ™ SYBR Green Supermix (BioRad Laboratories). .. The efficiency of the amplification reaction for each gene was calculated by running a standard curve of serially diluted cDNA sample, and the specificity of the amplification products was checked by analysis of the melting curve.

Article Title: Brachypodium distachyon as a model system for studies of copper transport in cereal crops
Article Snippet: Quantitative real-time (qRT)-PCR analysis Prior to qRT-PCR analysis, primers (Supplemental Table ) and cDNA concentrations were optimized to reach a qRT-PCR amplification efficiency of 100 ± 10%. .. Two microliters of 10-fold diluted cDNA was used as a template in a total reaction volume of 10 μl containing 500 nM of each PCR primer, 50 mM KCl, 20 mM Tris-HCI, pH 8.4, 0.2 mM dNTPs, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (BioRad), containing 3 mM MgCl2 , SYBR Green I, 20 nM fluoresceine, and stabilizers.

Article Title: Different skeletal effects of the peroxisome proliferator activated receptor (PPAR)? agonist fenofibrate and the PPAR? agonist pioglitazone
Article Snippet: .. The 2× iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad, Oslo, Norway). cDNA samples were analyzed both for the genes of interest and reference genes (β-actin) The amplification program consisted of a preincubation step for denaturation of the template cDNA (5 min, 95°C), followed by 40 cycles consisting of a denaturation step (30 s, 95°C), an annealing step (30 s, 60°C) and an extension step (30 s, 72°C). .. The Ct value, defined as the number of cycles required to produce a detectable product above background fluorescence, was measured for each sample, and arbitrary units were calculated using standard curves that consisted of serial dilutions of cDNA from a pool of samples or controls containing the highest amounts of the specific gene analyzed.

Article Title: Discovery of Single Nucleotide Polymorphisms in Complex Genomes Using SGSautoSNP
Article Snippet: .. PCR amplification of the 40 loci was performed using primers designed to conserved sequence surrounding the SNPs ( ) in a 20 μL reaction volume containing 1 × iTaq PCR buffer (containing 100 mM Tris-HCl and 500 mM KCl, pH 8.3) (Bio-Rad), 200 μM each dNTP (Bio-Rad), 0.5 μM each primer, 1.5 U iTaq DNA polymerase (Bio-Rad), RNase and DNase free water (Gibco) and 60 ng DNA. .. Final extension was performed at 72 °C for 10 min. Gel electrophoresis on a 1% (w/v) agarose gel in 1 × TAE buffer [ ] containing ethidium bromide resolved products, which were excised and purified using a silica method based on [ ].

Article Title: Cholesterol Defect Is Marked across Multiple Rodent Models of Huntington's Disease and Is Manifest in Astrocytes
Article Snippet: All reactions were performed in a total volume of 25 μl containing 2.5 ng of cDNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m forward and reverse primers. .. Amplification cycles consisted of an initial denaturing cycle at 95°C for 3 min, followed by 45 cycles of 30 s at 95°C, 30 s at 59°C for hmgcr and β-actin, at 55°C for cyp51 and 7dhcr, and 30 s at 72°C. iCycler ABI 7500 fast (Applied Biosystems) was used to evaluate gene expression in RNA from YAC astrocytes.

Article Title: Arabidopsis Pollen Fertility Requires the Transcription Factors CITF1 and SPL7 That Regulate Copper Delivery to Anthers and Jasmonic Acid Synthesis [OPEN]
Article Snippet: Prior to RT-qPCR analysis, primer and cDNA concentrations were optimized to reach the target and normalizing gene amplification efficiency of 100% ± 10%. .. Two microliters of 15-fold diluted cDNA was used as a template for RT-qPCR in a total volume of 15 μL containing a 500 nM concentration of each PCR primer, 50 mM KCl, 20 mM Tris-HCl, pH 8.4, 0.2 mM of each dNTP, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (Bio-Rad).

Article Title: Ameloblastin upstream region contains structural elements regulating transcriptional activity in a stromal cell line derived from bone marrow
Article Snippet: The 2X iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad). cDNA samples of M2-10B4 cells were analyzed for the mouse Ambn gene using the forward primer 5′-GCGTTTCCAAGAGCCCTGATAAC-3′ and the reverse primer 5′-AAGAAGCAGTGTCACATTTCCTGG-3′, resulting in a 366-bp product, and transfected M2-10B4 cells were analyzed for the LacZ gene using 5′-GCATTTTCCGTGACGTCTCGT-3′ as the forward primer and 5′-AACTCGCCGCACATCTGAACT-3′ as the reverse primer, resulting in a product of 130 bp. .. The amplification program consisted of a preincubation step for denaturation of the template cDNA (3 min, 95°C), followed by 50 and 40 cycles for Ambn and LacZ , respectively, consisting of a denaturation step (15 s, 95°C), an annealing step (30 s, 54°C and 60°C, Ambn and LacZ , respectively), and an extension step (30 s, 72°C).

Quantitative RT-PCR:

Article Title: Human Fibrocytes Express Multiple Antigens Associated With Autoimmune Endocrine Diseases
Article Snippet: Paragraph title: RNA isolation and quantitative RT-PCR ... Real-time PCR was performed using iQ SYBR Green Supermix (Bio-Rad Laboratories; catalog number 170–8882) in a reaction mixture also containing iTaq DNA polymerase, and deoxynucleotide triphosphates on a CFX96 real-time PCR system (Bio-Rad Laboratories).

Article Title: Loss of E2F1 Extends Survival and Accelerates Oral Tumor Growth in HPV-Positive Mice
Article Snippet: Paragraph title: 3.4. Quantitative RT-PCR ... QPCR was carried out using iQ™ SYBR® Green Supermix with hot-start iTaq DNA polymerase and iQ™ 5 Multicolor Real-time QPCR Detection System Machine (Bio-rad).

Article Title: Impaired glucose transporter-1 degradation and increased glucose transport and oxidative stress in response to high glucose in chondrocytes from osteoarthritic versus normal human cartilage
Article Snippet: .. Quantitative real-time RT-PCR was performed with iTaq™ DNA polymerase using iQ™ SYBR Green Supermix (BioRad Laboratories). .. The efficiency of the amplification reaction for each gene was calculated by running a standard curve of serially diluted cDNA sample, and the specificity of the amplification products was checked by analysis of the melting curve.

Article Title: Brachypodium distachyon as a model system for studies of copper transport in cereal crops
Article Snippet: Paragraph title: Quantitative real-time (qRT)-PCR analysis ... Two microliters of 10-fold diluted cDNA was used as a template in a total reaction volume of 10 μl containing 500 nM of each PCR primer, 50 mM KCl, 20 mM Tris-HCI, pH 8.4, 0.2 mM dNTPs, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (BioRad), containing 3 mM MgCl2 , SYBR Green I, 20 nM fluoresceine, and stabilizers.

Article Title: Arabidopsis Pollen Fertility Requires the Transcription Factors CITF1 and SPL7 That Regulate Copper Delivery to Anthers and Jasmonic Acid Synthesis [OPEN]
Article Snippet: .. Two microliters of 15-fold diluted cDNA was used as a template for RT-qPCR in a total volume of 15 μL containing a 500 nM concentration of each PCR primer, 50 mM KCl, 20 mM Tris-HCl, pH 8.4, 0.2 mM of each dNTP, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (Bio-Rad). .. PCR was performed using the CFX96 real-time PCR system (Bio-Rad).

Article Title: Ameloblastin upstream region contains structural elements regulating transcriptional activity in a stromal cell line derived from bone marrow
Article Snippet: Paragraph title: Real-time RT-PCR ... The 2X iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad). cDNA samples of M2-10B4 cells were analyzed for the mouse Ambn gene using the forward primer 5′-GCGTTTCCAAGAGCCCTGATAAC-3′ and the reverse primer 5′-AAGAAGCAGTGTCACATTTCCTGG-3′, resulting in a 366-bp product, and transfected M2-10B4 cells were analyzed for the LacZ gene using 5′-GCATTTTCCGTGACGTCTCGT-3′ as the forward primer and 5′-AACTCGCCGCACATCTGAACT-3′ as the reverse primer, resulting in a product of 130 bp.

SYBR Green Assay:

Article Title: Human Fibrocytes Express Multiple Antigens Associated With Autoimmune Endocrine Diseases
Article Snippet: .. Real-time PCR was performed using iQ SYBR Green Supermix (Bio-Rad Laboratories; catalog number 170–8882) in a reaction mixture also containing iTaq DNA polymerase, and deoxynucleotide triphosphates on a CFX96 real-time PCR system (Bio-Rad Laboratories). ..

Article Title: Loss of E2F1 Extends Survival and Accelerates Oral Tumor Growth in HPV-Positive Mice
Article Snippet: .. QPCR was carried out using iQ™ SYBR® Green Supermix with hot-start iTaq DNA polymerase and iQ™ 5 Multicolor Real-time QPCR Detection System Machine (Bio-rad). ..

Article Title: Impaired glucose transporter-1 degradation and increased glucose transport and oxidative stress in response to high glucose in chondrocytes from osteoarthritic versus normal human cartilage
Article Snippet: .. Quantitative real-time RT-PCR was performed with iTaq™ DNA polymerase using iQ™ SYBR Green Supermix (BioRad Laboratories). .. The efficiency of the amplification reaction for each gene was calculated by running a standard curve of serially diluted cDNA sample, and the specificity of the amplification products was checked by analysis of the melting curve.

Article Title: Brachypodium distachyon as a model system for studies of copper transport in cereal crops
Article Snippet: .. Two microliters of 10-fold diluted cDNA was used as a template in a total reaction volume of 10 μl containing 500 nM of each PCR primer, 50 mM KCl, 20 mM Tris-HCI, pH 8.4, 0.2 mM dNTPs, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (BioRad), containing 3 mM MgCl2 , SYBR Green I, 20 nM fluoresceine, and stabilizers. .. PCR was carried out using the CFX96 Real-Time PCR system (BioRad).

Article Title: Different skeletal effects of the peroxisome proliferator activated receptor (PPAR)? agonist fenofibrate and the PPAR? agonist pioglitazone
Article Snippet: .. The 2× iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad, Oslo, Norway). cDNA samples were analyzed both for the genes of interest and reference genes (β-actin) The amplification program consisted of a preincubation step for denaturation of the template cDNA (5 min, 95°C), followed by 40 cycles consisting of a denaturation step (30 s, 95°C), an annealing step (30 s, 60°C) and an extension step (30 s, 72°C). .. The Ct value, defined as the number of cycles required to produce a detectable product above background fluorescence, was measured for each sample, and arbitrary units were calculated using standard curves that consisted of serial dilutions of cDNA from a pool of samples or controls containing the highest amounts of the specific gene analyzed.

Article Title: Cholesterol Defect Is Marked across Multiple Rodent Models of Huntington's Disease and Is Manifest in Astrocytes
Article Snippet: .. All reactions were performed in a total volume of 25 μl containing 2.5 ng of cDNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m forward and reverse primers. .. Amplification cycles consisted of an initial denaturing cycle at 95°C for 3 min, followed by 45 cycles of 30 s at 95°C, 30 s at 59°C for hmgcr and β-actin, at 55°C for cyp51 and 7dhcr, and 30 s at 72°C. iCycler ABI 7500 fast (Applied Biosystems) was used to evaluate gene expression in RNA from YAC astrocytes.

Article Title: Disruption of astrocyte-neuron cholesterol cross talk affects neuronal function in Huntington's disease
Article Snippet: .. All reactions were performed in a total volume of 15 μ l, containing 60 ng cDNA, 50 mM KCl, 20 mM Tris-HCl (pH 8.4), 0.2 mM dNTPs, 25 U/ml iTaq DNA polymerase, 3 mM MgCl2 , SYBR Green I, 10 nM fluorescein, stabilizers (iQ EVA Green Supermix; Bio-Rad), and 0.2 μ M forward and reverse primers. .. Fluorescence was quantified during the annealing step, primer specificity and product formation were confirmed by melting curve analysis (55–94 °C), and the amounts of target gene mRNA were normalized to β -actin as reference gene and by using the calibration curve method.

Article Title: Arabidopsis Pollen Fertility Requires the Transcription Factors CITF1 and SPL7 That Regulate Copper Delivery to Anthers and Jasmonic Acid Synthesis [OPEN]
Article Snippet: .. Two microliters of 15-fold diluted cDNA was used as a template for RT-qPCR in a total volume of 15 μL containing a 500 nM concentration of each PCR primer, 50 mM KCl, 20 mM Tris-HCl, pH 8.4, 0.2 mM of each dNTP, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (Bio-Rad). .. PCR was performed using the CFX96 real-time PCR system (Bio-Rad).

Article Title: Ameloblastin upstream region contains structural elements regulating transcriptional activity in a stromal cell line derived from bone marrow
Article Snippet: .. The 2X iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad). cDNA samples of M2-10B4 cells were analyzed for the mouse Ambn gene using the forward primer 5′-GCGTTTCCAAGAGCCCTGATAAC-3′ and the reverse primer 5′-AAGAAGCAGTGTCACATTTCCTGG-3′, resulting in a 366-bp product, and transfected M2-10B4 cells were analyzed for the LacZ gene using 5′-GCATTTTCCGTGACGTCTCGT-3′ as the forward primer and 5′-AACTCGCCGCACATCTGAACT-3′ as the reverse primer, resulting in a product of 130 bp. .. The amplification program consisted of a preincubation step for denaturation of the template cDNA (3 min, 95°C), followed by 50 and 40 cycles for Ambn and LacZ , respectively, consisting of a denaturation step (15 s, 95°C), an annealing step (30 s, 54°C and 60°C, Ambn and LacZ , respectively), and an extension step (30 s, 72°C).

Article Title: Widespread Disruption of Repressor Element-1 Silencing Transcription Factor/Neuron-Restrictive Silencer Factor Occupancy at Its Target Genes in Huntington's Disease
Article Snippet: .. The PCR was performed in a total volume of 20 μl containing equal amounts of input and immunoprecipitated DNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m of forward and reverse primers. ..

Expressing:

Article Title: Loss of E2F1 Extends Survival and Accelerates Oral Tumor Growth in HPV-Positive Mice
Article Snippet: QPCR was carried out using iQ™ SYBR® Green Supermix with hot-start iTaq DNA polymerase and iQ™ 5 Multicolor Real-time QPCR Detection System Machine (Bio-rad). .. Normalized fold expression was obtained using the –log2∆∆ CT formula.

Article Title: Impaired glucose transporter-1 degradation and increased glucose transport and oxidative stress in response to high glucose in chondrocytes from osteoarthritic versus normal human cartilage
Article Snippet: Quantitative real-time RT-PCR was performed with iTaq™ DNA polymerase using iQ™ SYBR Green Supermix (BioRad Laboratories). .. Gene expression changes were analyzed using the built-in iQ5 Optical system software version 2, which enables the analysis of the results with the Pfaffl method, a variation of the ΔΔCT method corrected for gene-specific efficiencies.

Article Title: Cholesterol Defect Is Marked across Multiple Rodent Models of Huntington's Disease and Is Manifest in Astrocytes
Article Snippet: Paragraph title: RNA isolation, retrotranscription, and real-time PCR for gene expression. ... All reactions were performed in a total volume of 25 μl containing 2.5 ng of cDNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m forward and reverse primers.

Article Title: Disruption of astrocyte-neuron cholesterol cross talk affects neuronal function in Huntington's disease
Article Snippet: Paragraph title: RNA isolation, retrotranscription, and real-time PCR for gene expression ... All reactions were performed in a total volume of 15 μ l, containing 60 ng cDNA, 50 mM KCl, 20 mM Tris-HCl (pH 8.4), 0.2 mM dNTPs, 25 U/ml iTaq DNA polymerase, 3 mM MgCl2 , SYBR Green I, 10 nM fluorescein, stabilizers (iQ EVA Green Supermix; Bio-Rad), and 0.2 μ M forward and reverse primers.

Modification:

Article Title: Evaluation of p16INK4α Hypermethylation from Liquid-based Pap Test Samples in Vietnamese Population
Article Snippet: Other pair recognized a sequence in which CpG sites were unmethylated (modified to UpG treatment). .. MSP assay was carried out in a total of 15 μl containing 3 μl bisulfite-modified template DNA, 0.75 unit iTaq DNA polymerase (Biorad), 0.5 μM each primer, 7.5 μl MyTaqTM Mix (Bioline).

Transfection:

Article Title: Ameloblastin upstream region contains structural elements regulating transcriptional activity in a stromal cell line derived from bone marrow
Article Snippet: .. The 2X iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad). cDNA samples of M2-10B4 cells were analyzed for the mouse Ambn gene using the forward primer 5′-GCGTTTCCAAGAGCCCTGATAAC-3′ and the reverse primer 5′-AAGAAGCAGTGTCACATTTCCTGG-3′, resulting in a 366-bp product, and transfected M2-10B4 cells were analyzed for the LacZ gene using 5′-GCATTTTCCGTGACGTCTCGT-3′ as the forward primer and 5′-AACTCGCCGCACATCTGAACT-3′ as the reverse primer, resulting in a product of 130 bp. .. The amplification program consisted of a preincubation step for denaturation of the template cDNA (3 min, 95°C), followed by 50 and 40 cycles for Ambn and LacZ , respectively, consisting of a denaturation step (15 s, 95°C), an annealing step (30 s, 54°C and 60°C, Ambn and LacZ , respectively), and an extension step (30 s, 72°C).

Concentration Assay:

Article Title: Arabidopsis Pollen Fertility Requires the Transcription Factors CITF1 and SPL7 That Regulate Copper Delivery to Anthers and Jasmonic Acid Synthesis [OPEN]
Article Snippet: .. Two microliters of 15-fold diluted cDNA was used as a template for RT-qPCR in a total volume of 15 μL containing a 500 nM concentration of each PCR primer, 50 mM KCl, 20 mM Tris-HCl, pH 8.4, 0.2 mM of each dNTP, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (Bio-Rad). .. PCR was performed using the CFX96 real-time PCR system (Bio-Rad).

Reverse Transcription Polymerase Chain Reaction:

Article Title: Different skeletal effects of the peroxisome proliferator activated receptor (PPAR)? agonist fenofibrate and the PPAR? agonist pioglitazone
Article Snippet: The 2× iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad, Oslo, Norway). cDNA samples were analyzed both for the genes of interest and reference genes (β-actin) The amplification program consisted of a preincubation step for denaturation of the template cDNA (5 min, 95°C), followed by 40 cycles consisting of a denaturation step (30 s, 95°C), an annealing step (30 s, 60°C) and an extension step (30 s, 72°C). .. Contamination by genomic DNA was ruled out by performing PCR analysis where RT-enzyme had been omitted in the RT reactions. β-actin RT-PCR was run in duplicates or triplets as control to monitor RNA integrity and to be used for normalization.

Generated:

Article Title: Human Fibrocytes Express Multiple Antigens Associated With Autoimmune Endocrine Diseases
Article Snippet: Cellular RNA was extracted using the Aurum total RNA minikit (Bio-Rad Laboratories; catalog number 732–6820) according to the manufacturer's protocol. cDNA was generated by reverse transcription using oligo(deoxythymidine) and SuperScript III reverse transcriptase (Invitrogen Inc; catalog number 205311). .. Real-time PCR was performed using iQ SYBR Green Supermix (Bio-Rad Laboratories; catalog number 170–8882) in a reaction mixture also containing iTaq DNA polymerase, and deoxynucleotide triphosphates on a CFX96 real-time PCR system (Bio-Rad Laboratories).

Article Title: Loss of E2F1 Extends Survival and Accelerates Oral Tumor Growth in HPV-Positive Mice
Article Snippet: The cDNA was generated using ProtoScript® First Strand cDNA Synthesis Kit (NEB, Ipswich, MA, USA) and subjected to qPCR using SYBR Green Supermix (Biorad, Hercules, CA, USA). .. QPCR was carried out using iQ™ SYBR® Green Supermix with hot-start iTaq DNA polymerase and iQ™ 5 Multicolor Real-time QPCR Detection System Machine (Bio-rad).

Sequencing:

Article Title: Discovery of Single Nucleotide Polymorphisms in Complex Genomes Using SGSautoSNP
Article Snippet: .. PCR amplification of the 40 loci was performed using primers designed to conserved sequence surrounding the SNPs ( ) in a 20 μL reaction volume containing 1 × iTaq PCR buffer (containing 100 mM Tris-HCl and 500 mM KCl, pH 8.3) (Bio-Rad), 200 μM each dNTP (Bio-Rad), 0.5 μM each primer, 1.5 U iTaq DNA polymerase (Bio-Rad), RNase and DNase free water (Gibco) and 60 ng DNA. .. Final extension was performed at 72 °C for 10 min. Gel electrophoresis on a 1% (w/v) agarose gel in 1 × TAE buffer [ ] containing ethidium bromide resolved products, which were excised and purified using a silica method based on [ ].

Article Title: Evaluation of p16INK4α Hypermethylation from Liquid-based Pap Test Samples in Vietnamese Population
Article Snippet: Other pair recognized a sequence in which CpG sites were unmethylated (modified to UpG treatment). .. MSP assay was carried out in a total of 15 μl containing 3 μl bisulfite-modified template DNA, 0.75 unit iTaq DNA polymerase (Biorad), 0.5 μM each primer, 7.5 μl MyTaqTM Mix (Bioline).

Binding Assay:

Article Title: Widespread Disruption of Repressor Element-1 Silencing Transcription Factor/Neuron-Restrictive Silencer Factor Occupancy at Its Target Genes in Huntington's Disease
Article Snippet: A parallel approach was used to analyze REST/NRSF binding at the human BDNF locus (GenBank accession number ) using primers for flanking sequences located 758 bp upstream and 431, 1222, 2279, and 19350 bp downstream to the RE1/NRSE (for details on primers positions, see a ). .. The PCR was performed in a total volume of 20 μl containing equal amounts of input and immunoprecipitated DNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m of forward and reverse primers.

Nucleic Acid Electrophoresis:

Article Title: Discovery of Single Nucleotide Polymorphisms in Complex Genomes Using SGSautoSNP
Article Snippet: PCR amplification of the 40 loci was performed using primers designed to conserved sequence surrounding the SNPs ( ) in a 20 μL reaction volume containing 1 × iTaq PCR buffer (containing 100 mM Tris-HCl and 500 mM KCl, pH 8.3) (Bio-Rad), 200 μM each dNTP (Bio-Rad), 0.5 μM each primer, 1.5 U iTaq DNA polymerase (Bio-Rad), RNase and DNase free water (Gibco) and 60 ng DNA. .. Final extension was performed at 72 °C for 10 min. Gel electrophoresis on a 1% (w/v) agarose gel in 1 × TAE buffer [ ] containing ethidium bromide resolved products, which were excised and purified using a silica method based on [ ].

Fluorescence:

Article Title: Different skeletal effects of the peroxisome proliferator activated receptor (PPAR)? agonist fenofibrate and the PPAR? agonist pioglitazone
Article Snippet: The 2× iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad, Oslo, Norway). cDNA samples were analyzed both for the genes of interest and reference genes (β-actin) The amplification program consisted of a preincubation step for denaturation of the template cDNA (5 min, 95°C), followed by 40 cycles consisting of a denaturation step (30 s, 95°C), an annealing step (30 s, 60°C) and an extension step (30 s, 72°C). .. The Ct value, defined as the number of cycles required to produce a detectable product above background fluorescence, was measured for each sample, and arbitrary units were calculated using standard curves that consisted of serial dilutions of cDNA from a pool of samples or controls containing the highest amounts of the specific gene analyzed.

Article Title: Disruption of astrocyte-neuron cholesterol cross talk affects neuronal function in Huntington's disease
Article Snippet: All reactions were performed in a total volume of 15 μ l, containing 60 ng cDNA, 50 mM KCl, 20 mM Tris-HCl (pH 8.4), 0.2 mM dNTPs, 25 U/ml iTaq DNA polymerase, 3 mM MgCl2 , SYBR Green I, 10 nM fluorescein, stabilizers (iQ EVA Green Supermix; Bio-Rad), and 0.2 μ M forward and reverse primers. .. Fluorescence was quantified during the annealing step, primer specificity and product formation were confirmed by melting curve analysis (55–94 °C), and the amounts of target gene mRNA were normalized to β -actin as reference gene and by using the calibration curve method.

Article Title: Ameloblastin upstream region contains structural elements regulating transcriptional activity in a stromal cell line derived from bone marrow
Article Snippet: The 2X iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad). cDNA samples of M2-10B4 cells were analyzed for the mouse Ambn gene using the forward primer 5′-GCGTTTCCAAGAGCCCTGATAAC-3′ and the reverse primer 5′-AAGAAGCAGTGTCACATTTCCTGG-3′, resulting in a 366-bp product, and transfected M2-10B4 cells were analyzed for the LacZ gene using 5′-GCATTTTCCGTGACGTCTCGT-3′ as the forward primer and 5′-AACTCGCCGCACATCTGAACT-3′ as the reverse primer, resulting in a product of 130 bp. .. To allow relative quantification after PCR, standard curves were constructed from the reactions for each target and for two housekeeping genes – glyceraldehyde-3-phosphate dehydrogenase ( Gapdh ) and β - actin – by plotting cycle threshold ( C t ) values, that is, the cycle number at which the fluorescence signal exceeds background vs. log cDNA dilution.

Methylation:

Article Title: Evaluation of p16INK4α Hypermethylation from Liquid-based Pap Test Samples in Vietnamese Population
Article Snippet: Paragraph title: Methylation-specific polymerase chain reaction ... MSP assay was carried out in a total of 15 μl containing 3 μl bisulfite-modified template DNA, 0.75 unit iTaq DNA polymerase (Biorad), 0.5 μM each primer, 7.5 μl MyTaqTM Mix (Bioline).

Isolation:

Article Title: Human Fibrocytes Express Multiple Antigens Associated With Autoimmune Endocrine Diseases
Article Snippet: Paragraph title: RNA isolation and quantitative RT-PCR ... Real-time PCR was performed using iQ SYBR Green Supermix (Bio-Rad Laboratories; catalog number 170–8882) in a reaction mixture also containing iTaq DNA polymerase, and deoxynucleotide triphosphates on a CFX96 real-time PCR system (Bio-Rad Laboratories).

Article Title: Loss of E2F1 Extends Survival and Accelerates Oral Tumor Growth in HPV-Positive Mice
Article Snippet: Total RNA was isolated using TRIzol (Invitrogen, Carlsbad, CA, USA), and treated with DNase I. .. QPCR was carried out using iQ™ SYBR® Green Supermix with hot-start iTaq DNA polymerase and iQ™ 5 Multicolor Real-time QPCR Detection System Machine (Bio-rad).

Article Title: Discovery of Single Nucleotide Polymorphisms in Complex Genomes Using SGSautoSNP
Article Snippet: Genomic DNA was isolated from the four wheat cultivars Drysdale, Gladius, Excalibur and RAC875, according to a protocol adapted from [ ]. .. PCR amplification of the 40 loci was performed using primers designed to conserved sequence surrounding the SNPs ( ) in a 20 μL reaction volume containing 1 × iTaq PCR buffer (containing 100 mM Tris-HCl and 500 mM KCl, pH 8.3) (Bio-Rad), 200 μM each dNTP (Bio-Rad), 0.5 μM each primer, 1.5 U iTaq DNA polymerase (Bio-Rad), RNase and DNase free water (Gibco) and 60 ng DNA.

Article Title: Cholesterol Defect Is Marked across Multiple Rodent Models of Huntington's Disease and Is Manifest in Astrocytes
Article Snippet: Paragraph title: RNA isolation, retrotranscription, and real-time PCR for gene expression. ... All reactions were performed in a total volume of 25 μl containing 2.5 ng of cDNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m forward and reverse primers.

Article Title: Disruption of astrocyte-neuron cholesterol cross talk affects neuronal function in Huntington's disease
Article Snippet: Paragraph title: RNA isolation, retrotranscription, and real-time PCR for gene expression ... All reactions were performed in a total volume of 15 μ l, containing 60 ng cDNA, 50 mM KCl, 20 mM Tris-HCl (pH 8.4), 0.2 mM dNTPs, 25 U/ml iTaq DNA polymerase, 3 mM MgCl2 , SYBR Green I, 10 nM fluorescein, stabilizers (iQ EVA Green Supermix; Bio-Rad), and 0.2 μ M forward and reverse primers.

Article Title: Arabidopsis Pollen Fertility Requires the Transcription Factors CITF1 and SPL7 That Regulate Copper Delivery to Anthers and Jasmonic Acid Synthesis [OPEN]
Article Snippet: Total RNA was isolated using TRIzol reagent (Invitrogen) according to the manufacturer’s instructions. .. Two microliters of 15-fold diluted cDNA was used as a template for RT-qPCR in a total volume of 15 μL containing a 500 nM concentration of each PCR primer, 50 mM KCl, 20 mM Tris-HCl, pH 8.4, 0.2 mM of each dNTP, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (Bio-Rad).

RNA Extraction:

Article Title: Impaired glucose transporter-1 degradation and increased glucose transport and oxidative stress in response to high glucose in chondrocytes from osteoarthritic versus normal human cartilage
Article Snippet: Paragraph title: Total RNA extraction and quantitative real-time RT-PCR ... Quantitative real-time RT-PCR was performed with iTaq™ DNA polymerase using iQ™ SYBR Green Supermix (BioRad Laboratories).

Purification:

Article Title: Discovery of Single Nucleotide Polymorphisms in Complex Genomes Using SGSautoSNP
Article Snippet: PCR amplification of the 40 loci was performed using primers designed to conserved sequence surrounding the SNPs ( ) in a 20 μL reaction volume containing 1 × iTaq PCR buffer (containing 100 mM Tris-HCl and 500 mM KCl, pH 8.3) (Bio-Rad), 200 μM each dNTP (Bio-Rad), 0.5 μM each primer, 1.5 U iTaq DNA polymerase (Bio-Rad), RNase and DNase free water (Gibco) and 60 ng DNA. .. Final extension was performed at 72 °C for 10 min. Gel electrophoresis on a 1% (w/v) agarose gel in 1 × TAE buffer [ ] containing ethidium bromide resolved products, which were excised and purified using a silica method based on [ ].

Polymerase Chain Reaction:

Article Title: Brachypodium distachyon as a model system for studies of copper transport in cereal crops
Article Snippet: .. Two microliters of 10-fold diluted cDNA was used as a template in a total reaction volume of 10 μl containing 500 nM of each PCR primer, 50 mM KCl, 20 mM Tris-HCI, pH 8.4, 0.2 mM dNTPs, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (BioRad), containing 3 mM MgCl2 , SYBR Green I, 20 nM fluoresceine, and stabilizers. .. PCR was carried out using the CFX96 Real-Time PCR system (BioRad).

Article Title: Different skeletal effects of the peroxisome proliferator activated receptor (PPAR)? agonist fenofibrate and the PPAR? agonist pioglitazone
Article Snippet: The 2× iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad, Oslo, Norway). cDNA samples were analyzed both for the genes of interest and reference genes (β-actin) The amplification program consisted of a preincubation step for denaturation of the template cDNA (5 min, 95°C), followed by 40 cycles consisting of a denaturation step (30 s, 95°C), an annealing step (30 s, 60°C) and an extension step (30 s, 72°C). .. Contamination by genomic DNA was ruled out by performing PCR analysis where RT-enzyme had been omitted in the RT reactions. β-actin RT-PCR was run in duplicates or triplets as control to monitor RNA integrity and to be used for normalization.

Article Title: Discovery of Single Nucleotide Polymorphisms in Complex Genomes Using SGSautoSNP
Article Snippet: .. PCR amplification of the 40 loci was performed using primers designed to conserved sequence surrounding the SNPs ( ) in a 20 μL reaction volume containing 1 × iTaq PCR buffer (containing 100 mM Tris-HCl and 500 mM KCl, pH 8.3) (Bio-Rad), 200 μM each dNTP (Bio-Rad), 0.5 μM each primer, 1.5 U iTaq DNA polymerase (Bio-Rad), RNase and DNase free water (Gibco) and 60 ng DNA. .. Final extension was performed at 72 °C for 10 min. Gel electrophoresis on a 1% (w/v) agarose gel in 1 × TAE buffer [ ] containing ethidium bromide resolved products, which were excised and purified using a silica method based on [ ].

Article Title: Cholesterol Defect Is Marked across Multiple Rodent Models of Huntington's Disease and Is Manifest in Astrocytes
Article Snippet: All reactions were performed in a total volume of 25 μl containing 2.5 ng of cDNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m forward and reverse primers. .. All reactions were performed in a total volume of 10 μl containing 2.5 ng of cDNA, SYBR Green I Dye, AmpliTaqGold DNA Polymerase, dNTPs, passive reference and optimized buffer components (Power SYBR Green PCR Master Mix, Applied Biosystems), and 0.2 μ m forward and reverse primers.

Article Title: Arabidopsis Pollen Fertility Requires the Transcription Factors CITF1 and SPL7 That Regulate Copper Delivery to Anthers and Jasmonic Acid Synthesis [OPEN]
Article Snippet: .. Two microliters of 15-fold diluted cDNA was used as a template for RT-qPCR in a total volume of 15 μL containing a 500 nM concentration of each PCR primer, 50 mM KCl, 20 mM Tris-HCl, pH 8.4, 0.2 mM of each dNTP, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (Bio-Rad). .. PCR was performed using the CFX96 real-time PCR system (Bio-Rad).

Article Title: Ameloblastin upstream region contains structural elements regulating transcriptional activity in a stromal cell line derived from bone marrow
Article Snippet: The 2X iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad). cDNA samples of M2-10B4 cells were analyzed for the mouse Ambn gene using the forward primer 5′-GCGTTTCCAAGAGCCCTGATAAC-3′ and the reverse primer 5′-AAGAAGCAGTGTCACATTTCCTGG-3′, resulting in a 366-bp product, and transfected M2-10B4 cells were analyzed for the LacZ gene using 5′-GCATTTTCCGTGACGTCTCGT-3′ as the forward primer and 5′-AACTCGCCGCACATCTGAACT-3′ as the reverse primer, resulting in a product of 130 bp. .. To allow relative quantification after PCR, standard curves were constructed from the reactions for each target and for two housekeeping genes – glyceraldehyde-3-phosphate dehydrogenase ( Gapdh ) and β - actin – by plotting cycle threshold ( C t ) values, that is, the cycle number at which the fluorescence signal exceeds background vs. log cDNA dilution.

Article Title: Evaluation of p16INK4α Hypermethylation from Liquid-based Pap Test Samples in Vietnamese Population
Article Snippet: Paragraph title: Methylation-specific polymerase chain reaction ... MSP assay was carried out in a total of 15 μl containing 3 μl bisulfite-modified template DNA, 0.75 unit iTaq DNA polymerase (Biorad), 0.5 μM each primer, 7.5 μl MyTaqTM Mix (Bioline).

Article Title: Widespread Disruption of Repressor Element-1 Silencing Transcription Factor/Neuron-Restrictive Silencer Factor Occupancy at Its Target Genes in Huntington's Disease
Article Snippet: .. The PCR was performed in a total volume of 20 μl containing equal amounts of input and immunoprecipitated DNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m of forward and reverse primers. ..

MSP Assay:

Article Title: Evaluation of p16INK4α Hypermethylation from Liquid-based Pap Test Samples in Vietnamese Population
Article Snippet: .. MSP assay was carried out in a total of 15 μl containing 3 μl bisulfite-modified template DNA, 0.75 unit iTaq DNA polymerase (Biorad), 0.5 μM each primer, 7.5 μl MyTaqTM Mix (Bioline). .. Thermal cycling was initiated at 95 °C for 5 min, followed by 40 cycles of denaturation at 95 °C for 30 sec, annealing at the X °C for 30 sec, extension at 72 °C for 30 sec, and a final extension at 72 °C for 10 min (Note: X °C was the specific annealing temperature for each specific methylated or unmethylated primer).

Construct:

Article Title: Ameloblastin upstream region contains structural elements regulating transcriptional activity in a stromal cell line derived from bone marrow
Article Snippet: The 2X iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad). cDNA samples of M2-10B4 cells were analyzed for the mouse Ambn gene using the forward primer 5′-GCGTTTCCAAGAGCCCTGATAAC-3′ and the reverse primer 5′-AAGAAGCAGTGTCACATTTCCTGG-3′, resulting in a 366-bp product, and transfected M2-10B4 cells were analyzed for the LacZ gene using 5′-GCATTTTCCGTGACGTCTCGT-3′ as the forward primer and 5′-AACTCGCCGCACATCTGAACT-3′ as the reverse primer, resulting in a product of 130 bp. .. To allow relative quantification after PCR, standard curves were constructed from the reactions for each target and for two housekeeping genes – glyceraldehyde-3-phosphate dehydrogenase ( Gapdh ) and β - actin – by plotting cycle threshold ( C t ) values, that is, the cycle number at which the fluorescence signal exceeds background vs. log cDNA dilution.

IA:

Article Title: Human Fibrocytes Express Multiple Antigens Associated With Autoimmune Endocrine Diseases
Article Snippet: Real-time PCR was performed using iQ SYBR Green Supermix (Bio-Rad Laboratories; catalog number 170–8882) in a reaction mixture also containing iTaq DNA polymerase, and deoxynucleotide triphosphates on a CFX96 real-time PCR system (Bio-Rad Laboratories). .. Primer sequences were as follows: GAPDH forward, 5′-TTGCCATCAATGACCCCTTCA-3′, reverse, 5′-CGCCCCACTTGATTTTGGA-3′; IA-2, Hs00160947_m1; ICA69, Hs00245256_m1*; IL-8 forward, 5-CTTCCTGAT-TTCTGCAGCT-3′ and reverse, 5′-CCACTCTCAATCACTCTCAG-3′; IL-6 forward, 5′-TGAGAAAGGAGACATGTAACAAGAGT-3′, and reverse, 5-TTGTTCCTCACTACTCTCAAATCTGT-3′; TNF-α forward, 5′-GTCTCCTACCAGACCAAG-3′ and reverse, 5′-CAAAGTAGACCTGCCCAGACTC-3′.

Chromatin Immunoprecipitation:

Article Title: Impaired glucose transporter-1 degradation and increased glucose transport and oxidative stress in response to high glucose in chondrocytes from osteoarthritic versus normal human cartilage
Article Snippet: Total RNA extraction and quantitative real-time RT-PCR Total RNA was extracted with TRIzol (Invitrogen, Paisley, UK), analyzed using Experion RNA StdSens Chip (Bio-Rad Laboratories) and quantified in a NanoDrop ND-1000 Spectrophotometer (NanoDrop Technologies, Inc., Wilmington, DE, USA) at 260 nm. .. Quantitative real-time RT-PCR was performed with iTaq™ DNA polymerase using iQ™ SYBR Green Supermix (BioRad Laboratories).

Article Title: Widespread Disruption of Repressor Element-1 Silencing Transcription Factor/Neuron-Restrictive Silencer Factor Occupancy at Its Target Genes in Huntington's Disease
Article Snippet: Paragraph title: ChIP Scanning Assay. ... The PCR was performed in a total volume of 20 μl containing equal amounts of input and immunoprecipitated DNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m of forward and reverse primers.

Software:

Article Title: Impaired glucose transporter-1 degradation and increased glucose transport and oxidative stress in response to high glucose in chondrocytes from osteoarthritic versus normal human cartilage
Article Snippet: Specific sets of primers for GLUT-1 and endogenous control genes were designed using Beacon Designer software (PREMIER Biosoft International, Palo Alto, CA, USA). .. Quantitative real-time RT-PCR was performed with iTaq™ DNA polymerase using iQ™ SYBR Green Supermix (BioRad Laboratories).

Article Title: Brachypodium distachyon as a model system for studies of copper transport in cereal crops
Article Snippet: Two microliters of 10-fold diluted cDNA was used as a template in a total reaction volume of 10 μl containing 500 nM of each PCR primer, 50 mM KCl, 20 mM Tris-HCI, pH 8.4, 0.2 mM dNTPs, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (BioRad), containing 3 mM MgCl2 , SYBR Green I, 20 nM fluoresceine, and stabilizers. .. Data were analyzed using the CFX Manager Software, version 1.5 (BioRad).

Real-time Polymerase Chain Reaction:

Article Title: Human Fibrocytes Express Multiple Antigens Associated With Autoimmune Endocrine Diseases
Article Snippet: .. Real-time PCR was performed using iQ SYBR Green Supermix (Bio-Rad Laboratories; catalog number 170–8882) in a reaction mixture also containing iTaq DNA polymerase, and deoxynucleotide triphosphates on a CFX96 real-time PCR system (Bio-Rad Laboratories). ..

Article Title: Loss of E2F1 Extends Survival and Accelerates Oral Tumor Growth in HPV-Positive Mice
Article Snippet: .. QPCR was carried out using iQ™ SYBR® Green Supermix with hot-start iTaq DNA polymerase and iQ™ 5 Multicolor Real-time QPCR Detection System Machine (Bio-rad). ..

Article Title: Brachypodium distachyon as a model system for studies of copper transport in cereal crops
Article Snippet: Two microliters of 10-fold diluted cDNA was used as a template in a total reaction volume of 10 μl containing 500 nM of each PCR primer, 50 mM KCl, 20 mM Tris-HCI, pH 8.4, 0.2 mM dNTPs, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (BioRad), containing 3 mM MgCl2 , SYBR Green I, 20 nM fluoresceine, and stabilizers. .. PCR was carried out using the CFX96 Real-Time PCR system (BioRad).

Article Title: Different skeletal effects of the peroxisome proliferator activated receptor (PPAR)? agonist fenofibrate and the PPAR? agonist pioglitazone
Article Snippet: Paragraph title: Real time PCR quantification ... The 2× iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad, Oslo, Norway). cDNA samples were analyzed both for the genes of interest and reference genes (β-actin) The amplification program consisted of a preincubation step for denaturation of the template cDNA (5 min, 95°C), followed by 40 cycles consisting of a denaturation step (30 s, 95°C), an annealing step (30 s, 60°C) and an extension step (30 s, 72°C).

Article Title: Cholesterol Defect Is Marked across Multiple Rodent Models of Huntington's Disease and Is Manifest in Astrocytes
Article Snippet: Paragraph title: RNA isolation, retrotranscription, and real-time PCR for gene expression. ... All reactions were performed in a total volume of 25 μl containing 2.5 ng of cDNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m forward and reverse primers.

Article Title: Disruption of astrocyte-neuron cholesterol cross talk affects neuronal function in Huntington's disease
Article Snippet: Paragraph title: RNA isolation, retrotranscription, and real-time PCR for gene expression ... All reactions were performed in a total volume of 15 μ l, containing 60 ng cDNA, 50 mM KCl, 20 mM Tris-HCl (pH 8.4), 0.2 mM dNTPs, 25 U/ml iTaq DNA polymerase, 3 mM MgCl2 , SYBR Green I, 10 nM fluorescein, stabilizers (iQ EVA Green Supermix; Bio-Rad), and 0.2 μ M forward and reverse primers.

Article Title: Arabidopsis Pollen Fertility Requires the Transcription Factors CITF1 and SPL7 That Regulate Copper Delivery to Anthers and Jasmonic Acid Synthesis [OPEN]
Article Snippet: The AffinityScript QPCR cDNA synthesis kit (Agilent Technologies) was used for producing cDNA for subsequent RT-qPCR analyses. .. Two microliters of 15-fold diluted cDNA was used as a template for RT-qPCR in a total volume of 15 μL containing a 500 nM concentration of each PCR primer, 50 mM KCl, 20 mM Tris-HCl, pH 8.4, 0.2 mM of each dNTP, and 1.25 units of iTaq DNA polymerase in iQ SYBR Green Supermix (Bio-Rad).

Article Title: Widespread Disruption of Repressor Element-1 Silencing Transcription Factor/Neuron-Restrictive Silencer Factor Occupancy at Its Target Genes in Huntington's Disease
Article Snippet: The PCR was performed in a total volume of 20 μl containing equal amounts of input and immunoprecipitated DNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m of forward and reverse primers. .. DNA from the same chromatin immunoprecipitation underwent quantitative real-time PCR for β-actin, a gene not regulated by REST/NRSF and not proximal to any RE1/NRSEs.

Negative Control:

Article Title: Ameloblastin upstream region contains structural elements regulating transcriptional activity in a stromal cell line derived from bone marrow
Article Snippet: The 2X iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad). cDNA samples of M2-10B4 cells were analyzed for the mouse Ambn gene using the forward primer 5′-GCGTTTCCAAGAGCCCTGATAAC-3′ and the reverse primer 5′-AAGAAGCAGTGTCACATTTCCTGG-3′, resulting in a 366-bp product, and transfected M2-10B4 cells were analyzed for the LacZ gene using 5′-GCATTTTCCGTGACGTCTCGT-3′ as the forward primer and 5′-AACTCGCCGCACATCTGAACT-3′ as the reverse primer, resulting in a product of 130 bp. .. A negative control without cDNA template was run in each assay.

Agarose Gel Electrophoresis:

Article Title: Different skeletal effects of the peroxisome proliferator activated receptor (PPAR)? agonist fenofibrate and the PPAR? agonist pioglitazone
Article Snippet: The 2× iQ SYBR Green Supermix was based on iTaq DNA polymerase (Bio-Rad, Oslo, Norway). cDNA samples were analyzed both for the genes of interest and reference genes (β-actin) The amplification program consisted of a preincubation step for denaturation of the template cDNA (5 min, 95°C), followed by 40 cycles consisting of a denaturation step (30 s, 95°C), an annealing step (30 s, 60°C) and an extension step (30 s, 72°C). .. Specificity of each primer pair was confirmed by melting curve analysis and agarose gel electrophoresis, and PCR products were sequenced for product confirmation.

Article Title: Discovery of Single Nucleotide Polymorphisms in Complex Genomes Using SGSautoSNP
Article Snippet: PCR amplification of the 40 loci was performed using primers designed to conserved sequence surrounding the SNPs ( ) in a 20 μL reaction volume containing 1 × iTaq PCR buffer (containing 100 mM Tris-HCl and 500 mM KCl, pH 8.3) (Bio-Rad), 200 μM each dNTP (Bio-Rad), 0.5 μM each primer, 1.5 U iTaq DNA polymerase (Bio-Rad), RNase and DNase free water (Gibco) and 60 ng DNA. .. Final extension was performed at 72 °C for 10 min. Gel electrophoresis on a 1% (w/v) agarose gel in 1 × TAE buffer [ ] containing ethidium bromide resolved products, which were excised and purified using a silica method based on [ ].

Article Title: Evaluation of p16INK4α Hypermethylation from Liquid-based Pap Test Samples in Vietnamese Population
Article Snippet: MSP assay was carried out in a total of 15 μl containing 3 μl bisulfite-modified template DNA, 0.75 unit iTaq DNA polymerase (Biorad), 0.5 μM each primer, 7.5 μl MyTaqTM Mix (Bioline). .. The PCR products were run on 2% agarose gel with visualized by ethidium bromide staining.

Size-exclusion Chromatography:

Article Title: Evaluation of p16INK4α Hypermethylation from Liquid-based Pap Test Samples in Vietnamese Population
Article Snippet: MSP assay was carried out in a total of 15 μl containing 3 μl bisulfite-modified template DNA, 0.75 unit iTaq DNA polymerase (Biorad), 0.5 μM each primer, 7.5 μl MyTaqTM Mix (Bioline). .. Thermal cycling was initiated at 95 °C for 5 min, followed by 40 cycles of denaturation at 95 °C for 30 sec, annealing at the X °C for 30 sec, extension at 72 °C for 30 sec, and a final extension at 72 °C for 10 min (Note: X °C was the specific annealing temperature for each specific methylated or unmethylated primer).

Spectrophotometry:

Article Title: Impaired glucose transporter-1 degradation and increased glucose transport and oxidative stress in response to high glucose in chondrocytes from osteoarthritic versus normal human cartilage
Article Snippet: Total RNA extraction and quantitative real-time RT-PCR Total RNA was extracted with TRIzol (Invitrogen, Paisley, UK), analyzed using Experion RNA StdSens Chip (Bio-Rad Laboratories) and quantified in a NanoDrop ND-1000 Spectrophotometer (NanoDrop Technologies, Inc., Wilmington, DE, USA) at 260 nm. .. Quantitative real-time RT-PCR was performed with iTaq™ DNA polymerase using iQ™ SYBR Green Supermix (BioRad Laboratories).

Immunoprecipitation:

Article Title: Widespread Disruption of Repressor Element-1 Silencing Transcription Factor/Neuron-Restrictive Silencer Factor Occupancy at Its Target Genes in Huntington's Disease
Article Snippet: .. The PCR was performed in a total volume of 20 μl containing equal amounts of input and immunoprecipitated DNA, 50 m m KCl, 20 m m Tris-HCl, pH 8.4, 0.2 m m dNTPs, 25 U/ml iTaq DNA polymerase, 3 m m MgCl2 , SYBR Green I, 10 n m fluorescein, stabilizers (iQ SYBR Green Supermix; Bio-Rad), and 0.2 μ m of forward and reverse primers. ..

Staining:

Article Title: Evaluation of p16INK4α Hypermethylation from Liquid-based Pap Test Samples in Vietnamese Population
Article Snippet: MSP assay was carried out in a total of 15 μl containing 3 μl bisulfite-modified template DNA, 0.75 unit iTaq DNA polymerase (Biorad), 0.5 μM each primer, 7.5 μl MyTaqTM Mix (Bioline). .. The PCR products were run on 2% agarose gel with visualized by ethidium bromide staining.

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  • 88
    Bio-Rad i707l mutant cs3c klentaq1 dna polymerase
    A comparison of the <t>I707L</t> <t>KlenTaq1</t> <t>DNA</t> polymerase structures from MD simulations relative to crystal structures. (A) An overlay of the final MD structure (ice blue ribbons) with the I707L crystal structure (cyan ribbons). (B) An overlay of the final MD structure (ice blue ribbons) with the wild-type crystal structure (yellow ribbons). (C) A depiction of the active site from the simulated I707L mutant showing the four-residue pi-stacking interaction occurring between Phe667 on the O-helix of DNA polymerase, the base of C n-1 on the DNA primer strand, and the bases of G n –1 and G n from the template strand. The residue numbers correspond to the original 4KTQ.pdb. (D) A comparison of the I707L mutant binary complex (cyan) with the final structure of a molecular dynamics simulation of the I707 mutant binary complex at 2.2 μs (pink).
    I707l Mutant Cs3c Klentaq1 Dna Polymerase, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/i707l mutant cs3c klentaq1 dna polymerase/product/Bio-Rad
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    96
    Bio-Rad taq dna polymerase
    Superimposition of the thumb domains of <t>Taq</t> <t>DNA</t> polymerase (blue) with T7 DNA polymerase (pink). The arrows indicate the site of insertion of the T3 TBD (yellow). The primary amino acid sequence of Taq DNA polymerase from residue 470–507 is indicated below (blue) with the sequence of T3 TBD in yellow and the deleted region in red.
    Taq Dna Polymerase, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taq dna polymerase/product/Bio-Rad
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    86
    Bio-Rad iproof high fidelity dna polymerase pcr reaction mix
    Detection of VspC, VspQ, VspF1, and VspF2 expression by <t>RT-PCR</t> . Lane 1: V. spinosum RNA in the presence of reverse transcriptase and <t>DNA</t> polymerase; Lane 2: water (PCR negative control); Lane 3: V. spinosum RNA in the absence of reverse transcriptase and presence of DNA polymerase; Lane 4: V. spinosum DNA in the absence of reverse transcriptase and presence of DNA polymerase.
    Iproof High Fidelity Dna Polymerase Pcr Reaction Mix, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad hsv 1 dna polymerase plasmid
    Frequency of <t>HSV-1</t> <t>DNA</t> shedding over the course of the 30-day study (positive swabs/total swabs). Overall, 49 (98%) of the 50 subjects shed HSV DNA at least once. Only subject 7 did not shed virus in any of the samples. All 50 subjects are represented
    Hsv 1 Dna Polymerase Plasmid, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    A comparison of the I707L KlenTaq1 DNA polymerase structures from MD simulations relative to crystal structures. (A) An overlay of the final MD structure (ice blue ribbons) with the I707L crystal structure (cyan ribbons). (B) An overlay of the final MD structure (ice blue ribbons) with the wild-type crystal structure (yellow ribbons). (C) A depiction of the active site from the simulated I707L mutant showing the four-residue pi-stacking interaction occurring between Phe667 on the O-helix of DNA polymerase, the base of C n-1 on the DNA primer strand, and the bases of G n –1 and G n from the template strand. The residue numbers correspond to the original 4KTQ.pdb. (D) A comparison of the I707L mutant binary complex (cyan) with the final structure of a molecular dynamics simulation of the I707 mutant binary complex at 2.2 μs (pink).

    Journal: Biochemistry

    Article Title: A Conservative Isoleucine to Leucine Mutation Causes Major Rearrangements and Cold Sensitivity in KlenTaq1 DNA Polymerase

    doi: 10.1021/bi501198f

    Figure Lengend Snippet: A comparison of the I707L KlenTaq1 DNA polymerase structures from MD simulations relative to crystal structures. (A) An overlay of the final MD structure (ice blue ribbons) with the I707L crystal structure (cyan ribbons). (B) An overlay of the final MD structure (ice blue ribbons) with the wild-type crystal structure (yellow ribbons). (C) A depiction of the active site from the simulated I707L mutant showing the four-residue pi-stacking interaction occurring between Phe667 on the O-helix of DNA polymerase, the base of C n-1 on the DNA primer strand, and the bases of G n –1 and G n from the template strand. The residue numbers correspond to the original 4KTQ.pdb. (D) A comparison of the I707L mutant binary complex (cyan) with the final structure of a molecular dynamics simulation of the I707 mutant binary complex at 2.2 μs (pink).

    Article Snippet: Materials The wild-type and I707L mutant (Cs3C) KlenTaq1 DNA polymerase, which is an N-terminal deletion of 278 amino acids of Taq DNA polymerase, were produced and purified as previously described., Purified protein was dialyzed into buffer A (50 mM Tris-HCl, pH 7.5, 1 mM ethylenediaminetetraacetic acid, 6.5 mM 2-mercaptoethanol) and then bound to an equilibrated heparin agarose (Bio-Rad Laboratories, Hercules, CA) column to remove detergents for crystallography.

    Techniques: Mutagenesis

    Crystal structure of I707L KlenTaq1–DNA–dNTP ternary complex. (A) The crystal structure of the I707L mutant ternary complex (cyan) superimposed on the wild-type KlenTaq1 ternary complex (3KTQ; yellow). Residue 707 is connected to an active site divalent cation with a red dashed line corresponding to ∼24 Å long. (B) The 2 F o – F c map of the mutant ternary complex near the residue 707 mutation is shown at 1.0σ. (C) The active site, consisting of conserved aspartic acids and a ddCTP, contains two metal ions, one of which is associated with strong anomalous difference density (displayed at 4.0σ in magenta) around a divalent cation site. The manganese(II) atom is connected to nearby oxygen atoms with purple dashed lines corresponding to ∼2.1 Å long. The interatomic distances between the magnesium(II) atom and surrounding oxygen atoms are also ∼2.1 Å.

    Journal: Biochemistry

    Article Title: A Conservative Isoleucine to Leucine Mutation Causes Major Rearrangements and Cold Sensitivity in KlenTaq1 DNA Polymerase

    doi: 10.1021/bi501198f

    Figure Lengend Snippet: Crystal structure of I707L KlenTaq1–DNA–dNTP ternary complex. (A) The crystal structure of the I707L mutant ternary complex (cyan) superimposed on the wild-type KlenTaq1 ternary complex (3KTQ; yellow). Residue 707 is connected to an active site divalent cation with a red dashed line corresponding to ∼24 Å long. (B) The 2 F o – F c map of the mutant ternary complex near the residue 707 mutation is shown at 1.0σ. (C) The active site, consisting of conserved aspartic acids and a ddCTP, contains two metal ions, one of which is associated with strong anomalous difference density (displayed at 4.0σ in magenta) around a divalent cation site. The manganese(II) atom is connected to nearby oxygen atoms with purple dashed lines corresponding to ∼2.1 Å long. The interatomic distances between the magnesium(II) atom and surrounding oxygen atoms are also ∼2.1 Å.

    Article Snippet: Materials The wild-type and I707L mutant (Cs3C) KlenTaq1 DNA polymerase, which is an N-terminal deletion of 278 amino acids of Taq DNA polymerase, were produced and purified as previously described., Purified protein was dialyzed into buffer A (50 mM Tris-HCl, pH 7.5, 1 mM ethylenediaminetetraacetic acid, 6.5 mM 2-mercaptoethanol) and then bound to an equilibrated heparin agarose (Bio-Rad Laboratories, Hercules, CA) column to remove detergents for crystallography.

    Techniques: Mutagenesis

    Crystal structure of I707L KlenTaq1–DNA binary complex. (A) The crystal structure of the I707L mutant binary complex is colored by B-factor (ranging from 30 (blue) to 100 (red)). Portions of the fingers subdomain that could not be modeled are denoted by dashed lines (black, residues 637–660; blue, residues 673–699). (B) The fingers subdomain was more difficult to model in the binary complex (gray) compared to the ternary complex (cyan). Gray mesh, 2 F o – F c map of binary complex at 1.0σ. (C) Conformation of the DNA (colored ball and sticks) and O helix near the active site of I707L KlenTaq1 binary complex containing an AAA overhang (magenta) vs a TTT overhang (green). The DNA backbone is traced with a thick tube. The shift in the O helix angle is shown with an arrow. Nucleotides are numbered relative to the nucleotide to be copied ( n ). Manganese(II) ion bound to DNA is shown as a magenta ball.

    Journal: Biochemistry

    Article Title: A Conservative Isoleucine to Leucine Mutation Causes Major Rearrangements and Cold Sensitivity in KlenTaq1 DNA Polymerase

    doi: 10.1021/bi501198f

    Figure Lengend Snippet: Crystal structure of I707L KlenTaq1–DNA binary complex. (A) The crystal structure of the I707L mutant binary complex is colored by B-factor (ranging from 30 (blue) to 100 (red)). Portions of the fingers subdomain that could not be modeled are denoted by dashed lines (black, residues 637–660; blue, residues 673–699). (B) The fingers subdomain was more difficult to model in the binary complex (gray) compared to the ternary complex (cyan). Gray mesh, 2 F o – F c map of binary complex at 1.0σ. (C) Conformation of the DNA (colored ball and sticks) and O helix near the active site of I707L KlenTaq1 binary complex containing an AAA overhang (magenta) vs a TTT overhang (green). The DNA backbone is traced with a thick tube. The shift in the O helix angle is shown with an arrow. Nucleotides are numbered relative to the nucleotide to be copied ( n ). Manganese(II) ion bound to DNA is shown as a magenta ball.

    Article Snippet: Materials The wild-type and I707L mutant (Cs3C) KlenTaq1 DNA polymerase, which is an N-terminal deletion of 278 amino acids of Taq DNA polymerase, were produced and purified as previously described., Purified protein was dialyzed into buffer A (50 mM Tris-HCl, pH 7.5, 1 mM ethylenediaminetetraacetic acid, 6.5 mM 2-mercaptoethanol) and then bound to an equilibrated heparin agarose (Bio-Rad Laboratories, Hercules, CA) column to remove detergents for crystallography.

    Techniques: Mutagenesis

    Superimposition of the thumb domains of Taq DNA polymerase (blue) with T7 DNA polymerase (pink). The arrows indicate the site of insertion of the T3 TBD (yellow). The primary amino acid sequence of Taq DNA polymerase from residue 470–507 is indicated below (blue) with the sequence of T3 TBD in yellow and the deleted region in red.

    Journal: Nucleic Acids Research

    Article Title: Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase

    doi:

    Figure Lengend Snippet: Superimposition of the thumb domains of Taq DNA polymerase (blue) with T7 DNA polymerase (pink). The arrows indicate the site of insertion of the T3 TBD (yellow). The primary amino acid sequence of Taq DNA polymerase from residue 470–507 is indicated below (blue) with the sequence of T3 TBD in yellow and the deleted region in red.

    Article Snippet: Peak fractions contained a single polypeptide corresponding to the molecular weight for each of the Taq DNA polymerase (94 kDa), Taq /TDB (103 kDa) and Taq DNA polymerase (exo–) (68 kDa) and Taq /TDB(exo–) (76 kDa) variants, as evident by SDS–PAGE (10%) with Coomassie blue staining using molecular weight markers (Kaleidoscope Prestained Standards; Bio-Rad).

    Techniques: Sequencing

    Slippage chromatograms were obtained from PCR products amplified with either Taq DNA polymerase or Taq DNA pol/TBD. One primer was labeled with 6-FAM fluorophore and the PCR product was digested with EcoRI. The DNA was gel purified and slippage polymorphisms detected using an automated DNA sequencer (model 377; Applied Biosystems) and GENESCAN 672 software. The result is one representative of three experiments.

    Journal: Nucleic Acids Research

    Article Title: Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase

    doi:

    Figure Lengend Snippet: Slippage chromatograms were obtained from PCR products amplified with either Taq DNA polymerase or Taq DNA pol/TBD. One primer was labeled with 6-FAM fluorophore and the PCR product was digested with EcoRI. The DNA was gel purified and slippage polymorphisms detected using an automated DNA sequencer (model 377; Applied Biosystems) and GENESCAN 672 software. The result is one representative of three experiments.

    Article Snippet: Peak fractions contained a single polypeptide corresponding to the molecular weight for each of the Taq DNA polymerase (94 kDa), Taq /TDB (103 kDa) and Taq DNA polymerase (exo–) (68 kDa) and Taq /TDB(exo–) (76 kDa) variants, as evident by SDS–PAGE (10%) with Coomassie blue staining using molecular weight markers (Kaleidoscope Prestained Standards; Bio-Rad).

    Techniques: Polymerase Chain Reaction, Amplification, Labeling, Purification, Software

    Streptavidin processivity assay. An immobilized single-stranded DNA molecule of 2000 nt in length was incubated in a reaction containing a primer hybridized to the 5′ end, and polymerase. Extension was initiated by the addition of dNTPs including [α- 32 P]dGTP, Mg 2+ and 0.8 mg/ml activated calf thymus DNA as described in Materials and Methods. Cleavage with restriction enzymes located 18, 96, 492, 1122 and 1898 nt, respectively, from the primer terminus only occurs if primer extension results in a double-stranded DNA substrate. Full extension with 5 U Promega Taq DNA polymerase in the absence of trap DNA allowed the percentage of primers extended to be determined.

    Journal: Nucleic Acids Research

    Article Title: Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase

    doi:

    Figure Lengend Snippet: Streptavidin processivity assay. An immobilized single-stranded DNA molecule of 2000 nt in length was incubated in a reaction containing a primer hybridized to the 5′ end, and polymerase. Extension was initiated by the addition of dNTPs including [α- 32 P]dGTP, Mg 2+ and 0.8 mg/ml activated calf thymus DNA as described in Materials and Methods. Cleavage with restriction enzymes located 18, 96, 492, 1122 and 1898 nt, respectively, from the primer terminus only occurs if primer extension results in a double-stranded DNA substrate. Full extension with 5 U Promega Taq DNA polymerase in the absence of trap DNA allowed the percentage of primers extended to be determined.

    Article Snippet: Peak fractions contained a single polypeptide corresponding to the molecular weight for each of the Taq DNA polymerase (94 kDa), Taq /TDB (103 kDa) and Taq DNA polymerase (exo–) (68 kDa) and Taq /TDB(exo–) (76 kDa) variants, as evident by SDS–PAGE (10%) with Coomassie blue staining using molecular weight markers (Kaleidoscope Prestained Standards; Bio-Rad).

    Techniques: Incubation

    The effect of thioredoxin on processivity of the hybrid Taq DNA pol/TBD. ( A ) Extension assays were performed with a molar excess of template corresponding to a primer/template ratio of 470 for Taq DNA polymerase and Taq DNA polymerase (exo–) and 67 for Taq DNA pol/TBD and Taq DNA pol/TBD(exo–). Different ratios for the enzymes were used to ensure equal activity was loaded on the gel. (+) 100 µM thioredoxin, (–) no thioredoxin. No enzyme control shows the labeled primer alone. ( B ) Extension assay showing the effect of increasing concentrations of thioredoxin and enzyme dilution for Taq DNA pol/TBD(exo–). For each thioredoxin concentration (0.2, 2 and 20 µM), three enzyme concentrations were used (56, 28 and 5.6 pM) corresponding to a primer/template ratio of 67, 134 and 670.

    Journal: Nucleic Acids Research

    Article Title: Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase

    doi:

    Figure Lengend Snippet: The effect of thioredoxin on processivity of the hybrid Taq DNA pol/TBD. ( A ) Extension assays were performed with a molar excess of template corresponding to a primer/template ratio of 470 for Taq DNA polymerase and Taq DNA polymerase (exo–) and 67 for Taq DNA pol/TBD and Taq DNA pol/TBD(exo–). Different ratios for the enzymes were used to ensure equal activity was loaded on the gel. (+) 100 µM thioredoxin, (–) no thioredoxin. No enzyme control shows the labeled primer alone. ( B ) Extension assay showing the effect of increasing concentrations of thioredoxin and enzyme dilution for Taq DNA pol/TBD(exo–). For each thioredoxin concentration (0.2, 2 and 20 µM), three enzyme concentrations were used (56, 28 and 5.6 pM) corresponding to a primer/template ratio of 67, 134 and 670.

    Article Snippet: Peak fractions contained a single polypeptide corresponding to the molecular weight for each of the Taq DNA polymerase (94 kDa), Taq /TDB (103 kDa) and Taq DNA polymerase (exo–) (68 kDa) and Taq /TDB(exo–) (76 kDa) variants, as evident by SDS–PAGE (10%) with Coomassie blue staining using molecular weight markers (Kaleidoscope Prestained Standards; Bio-Rad).

    Techniques: Activity Assay, Labeling, Concentration Assay

    Detection of VspC, VspQ, VspF1, and VspF2 expression by RT-PCR . Lane 1: V. spinosum RNA in the presence of reverse transcriptase and DNA polymerase; Lane 2: water (PCR negative control); Lane 3: V. spinosum RNA in the absence of reverse transcriptase and presence of DNA polymerase; Lane 4: V. spinosum DNA in the absence of reverse transcriptase and presence of DNA polymerase.

    Journal: Frontiers in Microbiology

    Article Title: Genomic and Experimental Evidence Suggests that Verrucomicrobium spinosum Interacts with Eukaryotes

    doi: 10.3389/fmicb.2011.00211

    Figure Lengend Snippet: Detection of VspC, VspQ, VspF1, and VspF2 expression by RT-PCR . Lane 1: V. spinosum RNA in the presence of reverse transcriptase and DNA polymerase; Lane 2: water (PCR negative control); Lane 3: V. spinosum RNA in the absence of reverse transcriptase and presence of DNA polymerase; Lane 4: V. spinosum DNA in the absence of reverse transcriptase and presence of DNA polymerase.

    Article Snippet: PCR was performed in a mixture of 1 × iProof High-Fidelity DNA polymerase PCR reaction mix (Bio-Rad), 25 μM each primer, and approximately 30 ng of V. spinosum gDNA in a final volume of 50 μl.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Negative Control

    Frequency of HSV-1 DNA shedding over the course of the 30-day study (positive swabs/total swabs). Overall, 49 (98%) of the 50 subjects shed HSV DNA at least once. Only subject 7 did not shed virus in any of the samples. All 50 subjects are represented

    Journal:

    Article Title: HSV-1 DNA in Tears and Saliva of Normal Adults

    doi: 10.1167/iovs.04-0614

    Figure Lengend Snippet: Frequency of HSV-1 DNA shedding over the course of the 30-day study (positive swabs/total swabs). Overall, 49 (98%) of the 50 subjects shed HSV DNA at least once. Only subject 7 did not shed virus in any of the samples. All 50 subjects are represented

    Article Snippet: Nine serial dilutions of a known concentration of an HSV-1 DNA polymerase plasmid were assayed in triplicate, and the baseline threshold was determined by the thermocycler system software (iCycler; Bio-Rad) to be 30.

    Techniques:

    Distribution of HSV-1 DNA copy numbers per 10 μL for 941 positive eye and 1020 positive mouth swabs.

    Journal:

    Article Title: HSV-1 DNA in Tears and Saliva of Normal Adults

    doi: 10.1167/iovs.04-0614

    Figure Lengend Snippet: Distribution of HSV-1 DNA copy numbers per 10 μL for 941 positive eye and 1020 positive mouth swabs.

    Article Snippet: Nine serial dilutions of a known concentration of an HSV-1 DNA polymerase plasmid were assayed in triplicate, and the baseline threshold was determined by the thermocycler system software (iCycler; Bio-Rad) to be 30.

    Techniques:

    Distribution of 2806 eye and 2723 mouth swab results obtained with HSV-1 DNA polymerase real-time PCR. Of the total swabs assayed, 941 (33.5%) eye and 1020 (37.5%) mouth swabs were positive.

    Journal:

    Article Title: HSV-1 DNA in Tears and Saliva of Normal Adults

    doi: 10.1167/iovs.04-0614

    Figure Lengend Snippet: Distribution of 2806 eye and 2723 mouth swab results obtained with HSV-1 DNA polymerase real-time PCR. Of the total swabs assayed, 941 (33.5%) eye and 1020 (37.5%) mouth swabs were positive.

    Article Snippet: Nine serial dilutions of a known concentration of an HSV-1 DNA polymerase plasmid were assayed in triplicate, and the baseline threshold was determined by the thermocycler system software (iCycler; Bio-Rad) to be 30.

    Techniques: Real-time Polymerase Chain Reaction

    Distribution of HSV-1 DNA copy numbers in tears from subjects 3, 16, and 28 for 30 consecutive days. AM samples were obtained before oral hygiene was performed. PM samples were obtained approximately 12 hours later.

    Journal:

    Article Title: HSV-1 DNA in Tears and Saliva of Normal Adults

    doi: 10.1167/iovs.04-0614

    Figure Lengend Snippet: Distribution of HSV-1 DNA copy numbers in tears from subjects 3, 16, and 28 for 30 consecutive days. AM samples were obtained before oral hygiene was performed. PM samples were obtained approximately 12 hours later.

    Article Snippet: Nine serial dilutions of a known concentration of an HSV-1 DNA polymerase plasmid were assayed in triplicate, and the baseline threshold was determined by the thermocycler system software (iCycler; Bio-Rad) to be 30.

    Techniques: