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    PCR Master Mix provides an efficient accurate and convenient way to amplify DNA from any template This pre mixed formulation saves time and reduces contamination due to a reduced number
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    Thermo Fisher pcr mixture
    Pcr Mixture, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 11182 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega pcr mixture
    Pcr Mixture, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 3256 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad pcr mixture
    Viral replication and blood clinical chemistry at 3 dpi. Outbred CD1 mice ( n = 3 per group) were challenged with a 1 × 10 3 -PFU dose (i.p.) of parental strain rZH501, phosphate-buffered saline (mock-infected controls), or rZH501 with the U795C (Y259H) or A3564G (R1182G) mutation. At 3 dpi, the mice were euthanized and whole blood, sera, or total RNA from livers or spleens was analyzed. (A) Virus titer determined by plaque assay. The detection limit (10 2 PFU/ml) is shown by a dotted line. (B and C) Viral RNA copy numbers per 1 μg of total RNA in the liver (B) and spleen (C) at 3 dpi were determined by droplet digital <t>PCR</t> using a TaqMan probe detecting the <t>RVFV</t> S segment at the N ORF. (D to F) VetScan comprehensive diagnostic profile (Abaxis) test results for ALT (D), ALP (E), and total bilirubin (F) using heparinized whole-blood samples obtained at 3 dpi. The bars in panels A to F represent means and standard errors. (G) Immunohistochemistry of livers from mice infected with the U795C [Y259H (Gn)] (left) or A3564G [R1182G (Gc)] (right) mutant. Arrows, localization of RVFV N antigens.
    Pcr Mixture, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 1531 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa pcr buffer
    Viral replication and blood clinical chemistry at 3 dpi. Outbred CD1 mice ( n = 3 per group) were challenged with a 1 × 10 3 -PFU dose (i.p.) of parental strain rZH501, phosphate-buffered saline (mock-infected controls), or rZH501 with the U795C (Y259H) or A3564G (R1182G) mutation. At 3 dpi, the mice were euthanized and whole blood, sera, or total RNA from livers or spleens was analyzed. (A) Virus titer determined by plaque assay. The detection limit (10 2 PFU/ml) is shown by a dotted line. (B and C) Viral RNA copy numbers per 1 μg of total RNA in the liver (B) and spleen (C) at 3 dpi were determined by droplet digital <t>PCR</t> using a TaqMan probe detecting the <t>RVFV</t> S segment at the N ORF. (D to F) VetScan comprehensive diagnostic profile (Abaxis) test results for ALT (D), ALP (E), and total bilirubin (F) using heparinized whole-blood samples obtained at 3 dpi. The bars in panels A to F represent means and standard errors. (G) Immunohistochemistry of livers from mice infected with the U795C [Y259H (Gn)] (left) or A3564G [R1182G (Gc)] (right) mutant. Arrows, localization of RVFV N antigens.
    Pcr Buffer, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 10602 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore pcr mixture
    Viral replication and blood clinical chemistry at 3 dpi. Outbred CD1 mice ( n = 3 per group) were challenged with a 1 × 10 3 -PFU dose (i.p.) of parental strain rZH501, phosphate-buffered saline (mock-infected controls), or rZH501 with the U795C (Y259H) or A3564G (R1182G) mutation. At 3 dpi, the mice were euthanized and whole blood, sera, or total RNA from livers or spleens was analyzed. (A) Virus titer determined by plaque assay. The detection limit (10 2 PFU/ml) is shown by a dotted line. (B and C) Viral RNA copy numbers per 1 μg of total RNA in the liver (B) and spleen (C) at 3 dpi were determined by droplet digital <t>PCR</t> using a TaqMan probe detecting the <t>RVFV</t> S segment at the N ORF. (D to F) VetScan comprehensive diagnostic profile (Abaxis) test results for ALT (D), ALP (E), and total bilirubin (F) using heparinized whole-blood samples obtained at 3 dpi. The bars in panels A to F represent means and standard errors. (G) Immunohistochemistry of livers from mice infected with the U795C [Y259H (Gn)] (left) or A3564G [R1182G (Gc)] (right) mutant. Arrows, localization of RVFV N antigens.
    Pcr Mixture, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 977 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad pcr reaction mixture
    An example showing the effect of sample matrix on <t>ddPCR</t> output <t>(copies/PCR).</t> A cooked chicken sample was spiked in pork summer sausage, beef hot dog or beef and pork salami matrices.
    Pcr Reaction Mixture, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 737 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher pcr reaction mixture
    An example showing the effect of sample matrix on <t>ddPCR</t> output <t>(copies/PCR).</t> A cooked chicken sample was spiked in pork summer sausage, beef hot dog or beef and pork salami matrices.
    Pcr Reaction Mixture, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 5390 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boehringer Mannheim pcr mixture
    Detection of the linear form of HIV-1 <t>DNA</t> using <t>LM-PCR.</t> The final product of the reverse transcription reaction is a linear genome with blunt ends. To detect linear products of the reverse transcription reaction, genomic DNA was isolated from infected cells and loaded into a ligation reaction mixture containing a vast molar excess of an asymmetric blunt linker. The ligation of linker onto the end of the genome was detected by PCR with an HIV-1-specific primer and one complementary to the linker. In some experiments, a heminested PCR approach was employed. Production of an HIV-1-specific amplicon was universally T4 ligase dependent.
    Pcr Mixture, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 498 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene pcr mixture
    <t>RT-PCR</t> analysis of ERα and β transcripts in the heart chambers of newborn (4-day-old) and adult (55-day-old) female rats. Total RNA (1 μg) was reverse-transcribed, and <t>cDNA</t> was exponentially amplified in the presence of gene-specific primers. ( A ) A PhosphorImager scan after separation of the radioactive PCR product by electrophoresis in 1.5% agarose. Representative bands obtained from three RNA samples of the right atrium (RA) and left ventricle (LV) are shown. ( B ) Bar graphs depict ERα mRNA ( Left ) and ERβ mRNA ( Right ) levels verified by 18S RNA product used as internal standard. The results were calculated as a ratio of the signal given by radioactive bands of ERα mRNA or ERβ mRNA to the corresponding 18S band. RNA samples from the whole heart of newborn (1-day-old) rats were included in each experiment and used as a standard (100%) for calibration of ERα mRNA or ERβ mRNA levels. Values are means ± SEM for the right atrium and left ventricle, as well as for the left atrium (LA) and right ventricle (RV). n = 6; §, P
    Pcr Mixture, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 522 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Toyobo pcr mixture
    Expression of survivin mRNA as assessed by <t>RT-PCR</t> in normal tissues, and oral cancer cell lines and primary oral cancer tissues . (A) Expression of survivin mRNA in oral cancer cell lines and primary oral cancer tissues from two patients. G3PDH expression was detected as an internal control. (B) Expression of survivin mRNA in normal adult tissues. 293T cells transfected with myc-tagged survivin <t>cDNA</t> (293T-survivin) was used as a positive control for survivin expression. G3PDH expression was detected as an internal control.
    Pcr Mixture, supplied by Toyobo, used in various techniques. Bioz Stars score: 92/100, based on 441 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher sybr green pcr master mixture
    Expression of survivin mRNA as assessed by <t>RT-PCR</t> in normal tissues, and oral cancer cell lines and primary oral cancer tissues . (A) Expression of survivin mRNA in oral cancer cell lines and primary oral cancer tissues from two patients. G3PDH expression was detected as an internal control. (B) Expression of survivin mRNA in normal adult tissues. 293T cells transfected with myc-tagged survivin <t>cDNA</t> (293T-survivin) was used as a positive control for survivin expression. G3PDH expression was detected as an internal control.
    Sybr Green Pcr Master Mixture, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 382 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman universal pcr master mixture
    Expression of survivin mRNA as assessed by <t>RT-PCR</t> in normal tissues, and oral cancer cell lines and primary oral cancer tissues . (A) Expression of survivin mRNA in oral cancer cell lines and primary oral cancer tissues from two patients. G3PDH expression was detected as an internal control. (B) Expression of survivin mRNA in normal adult tissues. 293T cells transfected with myc-tagged survivin <t>cDNA</t> (293T-survivin) was used as a positive control for survivin expression. G3PDH expression was detected as an internal control.
    Taqman Universal Pcr Master Mixture, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 401 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore redtaq readymix pcr reaction mixture
    Expression of survivin mRNA as assessed by <t>RT-PCR</t> in normal tissues, and oral cancer cell lines and primary oral cancer tissues . (A) Expression of survivin mRNA in oral cancer cell lines and primary oral cancer tissues from two patients. G3PDH expression was detected as an internal control. (B) Expression of survivin mRNA in normal adult tissues. 293T cells transfected with myc-tagged survivin <t>cDNA</t> (293T-survivin) was used as a positive control for survivin expression. G3PDH expression was detected as an internal control.
    Redtaq Readymix Pcr Reaction Mixture, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATUM pcr mixture
    Methylation-specific <t>PCR</t> results of some randomly selected sample patients with schizophrenia and control groups, analyzed with 2% agarose gel electrophoresis followed by ethidium bromide staining. A: Methylation pattern of some patients in CpG island-1 from BDNF gene. B: Methylation pattern of some controls in CpG island-1 from BDNF gene. C: Methylation pattern of some patients in CpG island-2 from BDNF gene. D: Methylation pattern of some controls in CpG island-2 from BDNF gene. PC: Positive control; M: methylated; U: unmethylated; 100M : 100 bp <t>DNA</t> ladder.
    Pcr Mixture, supplied by ATUM, used in various techniques. Bioz Stars score: 92/100, based on 224 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Toyobo pcr reaction mixture
    Effects of HPV16 E1 mutants on HPV18 replication. (A) Schematic representation of HPV16 E1 mutants. F, FLAG-tag; ND, N-terminal domain; DBD, DNA-binding domain; OD, oligomerization domain; HD, helicase domain. (B) C33A cells were transfected with 1 ng of the HPV18 gDNA together with the indicated amounts (ng) of the expression plasmids. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with <t>Dpn</t> I. The Dpn I-resistant HPV18 gDNA was quantified by real-time <t>PCR</t> and normalized to the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication with the HPV18 E1/E2 alone. Each bar represents the average of two independent experiments with the standard error of mean.
    Pcr Reaction Mixture, supplied by Toyobo, used in various techniques. Bioz Stars score: 92/100, based on 225 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher power sybr green pcr master mixture
    Effects of HPV16 E1 mutants on HPV18 replication. (A) Schematic representation of HPV16 E1 mutants. F, FLAG-tag; ND, N-terminal domain; DBD, DNA-binding domain; OD, oligomerization domain; HD, helicase domain. (B) C33A cells were transfected with 1 ng of the HPV18 gDNA together with the indicated amounts (ng) of the expression plasmids. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with <t>Dpn</t> I. The Dpn I-resistant HPV18 gDNA was quantified by real-time <t>PCR</t> and normalized to the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication with the HPV18 E1/E2 alone. Each bar represents the average of two independent experiments with the standard error of mean.
    Power Sybr Green Pcr Master Mixture, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 245 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher pcr master mixture
    Effects of HPV16 E1 mutants on HPV18 replication. (A) Schematic representation of HPV16 E1 mutants. F, FLAG-tag; ND, N-terminal domain; DBD, DNA-binding domain; OD, oligomerization domain; HD, helicase domain. (B) C33A cells were transfected with 1 ng of the HPV18 gDNA together with the indicated amounts (ng) of the expression plasmids. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with <t>Dpn</t> I. The Dpn I-resistant HPV18 gDNA was quantified by real-time <t>PCR</t> and normalized to the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication with the HPV18 E1/E2 alone. Each bar represents the average of two independent experiments with the standard error of mean.
    Pcr Master Mixture, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 208 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bioneer Corporation pcr mixture tube
    Effects of HPV16 E1 mutants on HPV18 replication. (A) Schematic representation of HPV16 E1 mutants. F, FLAG-tag; ND, N-terminal domain; DBD, DNA-binding domain; OD, oligomerization domain; HD, helicase domain. (B) C33A cells were transfected with 1 ng of the HPV18 gDNA together with the indicated amounts (ng) of the expression plasmids. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with <t>Dpn</t> I. The Dpn I-resistant HPV18 gDNA was quantified by real-time <t>PCR</t> and normalized to the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication with the HPV18 E1/E2 alone. Each bar represents the average of two independent experiments with the standard error of mean.
    Pcr Mixture Tube, supplied by Bioneer Corporation, used in various techniques. Bioz Stars score: 89/100, based on 256 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Kaneka Corp pcr mixture
    Standard curve generated from quantification cycle numbers of a 10-fold dilution series of an amplicon product of 86 bp cloned into a <t>pCR</t> 2.1 plasmid vector in DNase- and RNase-free water (≤5.00 to 5.00 × 10 5 copies/reaction). <t>qPCR</t> was
    Pcr Mixture, supplied by Kaneka Corp, used in various techniques. Bioz Stars score: 92/100, based on 169 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher pcr mixtures
    Standard curve generated from quantification cycle numbers of a 10-fold dilution series of an amplicon product of 86 bp cloned into a <t>pCR</t> 2.1 plasmid vector in DNase- and RNase-free water (≤5.00 to 5.00 × 10 5 copies/reaction). <t>qPCR</t> was
    Pcr Mixtures, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 2909 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    5 PRIME pcr mixture
    Genomic organization of MsHid . A. Schematic diagram of exon-intron structure. The horizontal boxes indicate exons (1 st exon, 657 bp; 2 nd exon, 145 bp; 3 rd exon, 205 bp; 4 th exon 249 bp). The locations of three introns are indicated by vertical lines that are accompanied with partial sequences (consensus 5′ and 3′ splicing junctions are indicated in reds). Arrows indicate <t>PCR</t> primers. A Hin P1 I site was used for inverse PCR. B. An agarose gel electrophoresis confirming introns. Primer sets are as follows: lane 1 and 2, 5′ORF-R2; lane 3, F3-Int1R; lane 4 and 5, RTF1-R1; lane 6, RTF1 and Int2R; lane 7 and 8, RTF2 and RTR2; lane 9, RTF2 and Int3R. Note that there is no PCR fragment in lane 1 and 7 due to large size of the 1 st and the 3 rd introns, respectively. However, PCR fragments from genomic <t>DNA</t> using the intron-derived primers match expected sizes (lane 3, 6, 9). (abbr.: G, genomic DNA template; C, cDNA template). C. Comparison of gene structure between DmHid (top) and MsHid (bottom). Exons are indicated by boxes, introns by lines. Numbers indicate nucleotide lengths of introns (asterisks for approximate values).
    Pcr Mixture, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 92/100, based on 189 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bioneer Corporation pcr mixture
    <t>PCR</t> assay for detection of bla VEB gene with product size: 600 bp. Lanes 3–17: Amplified products, Lanes 2 and 19: Positive control, Lanes 1 and 20: <t>DNA</t> ladder 100 bp, Lane 18: Negative control
    Pcr Mixture, supplied by Bioneer Corporation, used in various techniques. Bioz Stars score: 92/100, based on 219 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TransGen biotech co pcr mixture
    <t>PCR</t> assay for detection of bla VEB gene with product size: 600 bp. Lanes 3–17: Amplified products, Lanes 2 and 19: Positive control, Lanes 1 and 20: <t>DNA</t> ladder 100 bp, Lane 18: Negative control
    Pcr Mixture, supplied by TransGen biotech co, used in various techniques. Bioz Stars score: 92/100, based on 178 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Viral replication and blood clinical chemistry at 3 dpi. Outbred CD1 mice ( n = 3 per group) were challenged with a 1 × 10 3 -PFU dose (i.p.) of parental strain rZH501, phosphate-buffered saline (mock-infected controls), or rZH501 with the U795C (Y259H) or A3564G (R1182G) mutation. At 3 dpi, the mice were euthanized and whole blood, sera, or total RNA from livers or spleens was analyzed. (A) Virus titer determined by plaque assay. The detection limit (10 2 PFU/ml) is shown by a dotted line. (B and C) Viral RNA copy numbers per 1 μg of total RNA in the liver (B) and spleen (C) at 3 dpi were determined by droplet digital PCR using a TaqMan probe detecting the RVFV S segment at the N ORF. (D to F) VetScan comprehensive diagnostic profile (Abaxis) test results for ALT (D), ALP (E), and total bilirubin (F) using heparinized whole-blood samples obtained at 3 dpi. The bars in panels A to F represent means and standard errors. (G) Immunohistochemistry of livers from mice infected with the U795C [Y259H (Gn)] (left) or A3564G [R1182G (Gc)] (right) mutant. Arrows, localization of RVFV N antigens.

    Journal: Journal of Virology

    Article Title: Rift Valley Fever Virus MP-12 Vaccine Is Fully Attenuated by a Combination of Partial Attenuations in the S, M, and L Segments

    doi: 10.1128/JVI.00135-15

    Figure Lengend Snippet: Viral replication and blood clinical chemistry at 3 dpi. Outbred CD1 mice ( n = 3 per group) were challenged with a 1 × 10 3 -PFU dose (i.p.) of parental strain rZH501, phosphate-buffered saline (mock-infected controls), or rZH501 with the U795C (Y259H) or A3564G (R1182G) mutation. At 3 dpi, the mice were euthanized and whole blood, sera, or total RNA from livers or spleens was analyzed. (A) Virus titer determined by plaque assay. The detection limit (10 2 PFU/ml) is shown by a dotted line. (B and C) Viral RNA copy numbers per 1 μg of total RNA in the liver (B) and spleen (C) at 3 dpi were determined by droplet digital PCR using a TaqMan probe detecting the RVFV S segment at the N ORF. (D to F) VetScan comprehensive diagnostic profile (Abaxis) test results for ALT (D), ALP (E), and total bilirubin (F) using heparinized whole-blood samples obtained at 3 dpi. The bars in panels A to F represent means and standard errors. (G) Immunohistochemistry of livers from mice infected with the U795C [Y259H (Gn)] (left) or A3564G [R1182G (Gc)] (right) mutant. Arrows, localization of RVFV N antigens.

    Article Snippet: The total RNA concentration was measured by use of a Qubit 2.0 fluorometer (Life Technologies), and total RNA was used for first-strand cDNA synthesis by using an iScript cDNA synthesis kit (Bio-Rad), which contains a random hexamer and RNase H. cDNA derived from total RNA (25 or 250 ng), 250 nM TaqMan probe, 5′-HEX-CAG GCT TTG GTC GTC TTG AG-BHQ1-3′ (where HEX is hexachlorofluorescein and BHQ1 is black hole quencher 1), and 900 nM forward primer (5′-GGC TGG CTG GAC ATG-3′) and 900 nM reverse primer (5′-AGT GAC AGG AAG CCA CTC A-3′), which are specific for the RVFV N ORF, were added to the PCR mixture (25 μl), which also contained ddPCR supermix for probes (Bio-Rad).

    Techniques: Mouse Assay, Infection, Mutagenesis, Plaque Assay, Digital PCR, Diagnostic Assay, ALP Assay, Immunohistochemistry

    An example showing the effect of sample matrix on ddPCR output (copies/PCR). A cooked chicken sample was spiked in pork summer sausage, beef hot dog or beef and pork salami matrices.

    Journal: PLoS ONE

    Article Title: Droplet digital polymerase chain reaction (ddPCR) assays integrated with an internal control for quantification of bovine, porcine, chicken and turkey species in food and feed

    doi: 10.1371/journal.pone.0182872

    Figure Lengend Snippet: An example showing the effect of sample matrix on ddPCR output (copies/PCR). A cooked chicken sample was spiked in pork summer sausage, beef hot dog or beef and pork salami matrices.

    Article Snippet: The optimized PCR reaction mixture (25 μL/reaction) contained 1x ddPCR Supermix for Probe (Bio-Rad), 96 nM each of the primers and 64 nM probe for the animal target, 40 nM each of the primers and 32 nM probe for the internal control, 1700 copies of internal control plasmid DNA and 40–50 ng of template DNA.

    Techniques: Polymerase Chain Reaction

    Linear regression between ddPCR output (copies/PCR) and DNA amount (ng). DNA was extracted from fresh raw (A) beef, (B) pork, (C) chicken, and (D) turkey. Shown are results from two replicates in green and blue colors.

    Journal: PLoS ONE

    Article Title: Droplet digital polymerase chain reaction (ddPCR) assays integrated with an internal control for quantification of bovine, porcine, chicken and turkey species in food and feed

    doi: 10.1371/journal.pone.0182872

    Figure Lengend Snippet: Linear regression between ddPCR output (copies/PCR) and DNA amount (ng). DNA was extracted from fresh raw (A) beef, (B) pork, (C) chicken, and (D) turkey. Shown are results from two replicates in green and blue colors.

    Article Snippet: The optimized PCR reaction mixture (25 μL/reaction) contained 1x ddPCR Supermix for Probe (Bio-Rad), 96 nM each of the primers and 64 nM probe for the animal target, 40 nM each of the primers and 32 nM probe for the internal control, 1700 copies of internal control plasmid DNA and 40–50 ng of template DNA.

    Techniques: Polymerase Chain Reaction

    Linear regression between ddPCR output (copies/PCR) and target animal species concentration (% wt/wt) in fortified heat-processed samples. (A) beef in poultry meal, (B) pork in chicken, (C) chicken in pork, and (D) turkey in pork. Shown are results from four replicates in green, blue, purple and black.

    Journal: PLoS ONE

    Article Title: Droplet digital polymerase chain reaction (ddPCR) assays integrated with an internal control for quantification of bovine, porcine, chicken and turkey species in food and feed

    doi: 10.1371/journal.pone.0182872

    Figure Lengend Snippet: Linear regression between ddPCR output (copies/PCR) and target animal species concentration (% wt/wt) in fortified heat-processed samples. (A) beef in poultry meal, (B) pork in chicken, (C) chicken in pork, and (D) turkey in pork. Shown are results from four replicates in green, blue, purple and black.

    Article Snippet: The optimized PCR reaction mixture (25 μL/reaction) contained 1x ddPCR Supermix for Probe (Bio-Rad), 96 nM each of the primers and 64 nM probe for the animal target, 40 nM each of the primers and 32 nM probe for the internal control, 1700 copies of internal control plasmid DNA and 40–50 ng of template DNA.

    Techniques: Polymerase Chain Reaction, Concentration Assay

    Detection of the linear form of HIV-1 DNA using LM-PCR. The final product of the reverse transcription reaction is a linear genome with blunt ends. To detect linear products of the reverse transcription reaction, genomic DNA was isolated from infected cells and loaded into a ligation reaction mixture containing a vast molar excess of an asymmetric blunt linker. The ligation of linker onto the end of the genome was detected by PCR with an HIV-1-specific primer and one complementary to the linker. In some experiments, a heminested PCR approach was employed. Production of an HIV-1-specific amplicon was universally T4 ligase dependent.

    Journal: Journal of Virology

    Article Title: Molecular Characterization of Preintegration Latency in Human Immunodeficiency Virus Type 1 Infection

    doi: 10.1128/JVI.76.17.8518-8531.2002

    Figure Lengend Snippet: Detection of the linear form of HIV-1 DNA using LM-PCR. The final product of the reverse transcription reaction is a linear genome with blunt ends. To detect linear products of the reverse transcription reaction, genomic DNA was isolated from infected cells and loaded into a ligation reaction mixture containing a vast molar excess of an asymmetric blunt linker. The ligation of linker onto the end of the genome was detected by PCR with an HIV-1-specific primer and one complementary to the linker. In some experiments, a heminested PCR approach was employed. Production of an HIV-1-specific amplicon was universally T4 ligase dependent.

    Article Snippet: A mass of 200 ng of DNA was loaded into each PCR mixture, which contained 1 μM (each) primer, 200 μM deoxynucleoside triphosphates, 1.5 mM MgCl2 , and either 2.5 U of Expand High Fidelity Taq polymerase (Boehringer Mannheim) or 2.5 U of Platinum Taq (Life Technologies) in 1× proprietary buffer.

    Techniques: Polymerase Chain Reaction, Isolation, Infection, Ligation, Amplification

    Reverse transcription proceeds with protracted kinetics in highly purified resting T cells but can yield double-stranded HIV-1 DNA molecules of the appropriate length. Highly purified resting T cells were obtained from HIV-1-negative donors by cell sorting. Control MT-2 cells and resting CD4 + T cells were infected with HIV-1 IIIb at a multiplicity of infection of 1. At the indicated times, DNA was isolated and nested LM-PCR was performed to detect the U3 end of the linear HIV-1 genome. The LM-PCR involved an initial reaction of 15 cycles with outer LPCR-L and oligonucleotide 25t. Reaction products were diluted 1:4 in water, and a second 25-cycle PCR was performed using oligonucleotide 25t and LPCR-L. Reaction products were run on a 2% gel and Southern blotted with a U3-specific probe.

    Journal: Journal of Virology

    Article Title: Molecular Characterization of Preintegration Latency in Human Immunodeficiency Virus Type 1 Infection

    doi: 10.1128/JVI.76.17.8518-8531.2002

    Figure Lengend Snippet: Reverse transcription proceeds with protracted kinetics in highly purified resting T cells but can yield double-stranded HIV-1 DNA molecules of the appropriate length. Highly purified resting T cells were obtained from HIV-1-negative donors by cell sorting. Control MT-2 cells and resting CD4 + T cells were infected with HIV-1 IIIb at a multiplicity of infection of 1. At the indicated times, DNA was isolated and nested LM-PCR was performed to detect the U3 end of the linear HIV-1 genome. The LM-PCR involved an initial reaction of 15 cycles with outer LPCR-L and oligonucleotide 25t. Reaction products were diluted 1:4 in water, and a second 25-cycle PCR was performed using oligonucleotide 25t and LPCR-L. Reaction products were run on a 2% gel and Southern blotted with a U3-specific probe.

    Article Snippet: A mass of 200 ng of DNA was loaded into each PCR mixture, which contained 1 μM (each) primer, 200 μM deoxynucleoside triphosphates, 1.5 mM MgCl2 , and either 2.5 U of Expand High Fidelity Taq polymerase (Boehringer Mannheim) or 2.5 U of Platinum Taq (Life Technologies) in 1× proprietary buffer.

    Techniques: Purification, FACS, Infection, Isolation, Polymerase Chain Reaction

    Characterization of complete reverse transcripts cloned from resting T lymphocytes. (A) LM-PCR assay for complete reverse transcripts. The ligation of an asymmetric linker containing one-half of a Sca I site onto the terminus of a full-length HIV-1 genome creates a novel Sca I site, allowing rapid screening of cloned LM-PCR products. (B) Restriction analysis of cloned LM-PCR products obtained from resting CD4 + T cells. Highly purified resting CD4 + T cells were infected with HIV-1 IIIB. Three days after infection, cells were lysed and DNA was isolated for LM-PCR. Cloned LM-PCR products were restricted with Sca I and analyzed by Southern blotting using an LTR-specific probe. The analysis allowed the identification of blunt linear molecules with either the predicted U3 end (lanes 1, 2, 3, and 5) or clones bearing alterations that result in a change in the sequence of the most-terminal U3 nucleotides (lanes 4 and 6). Because the insert was cloned in either a forward or reverse orientation, inserts of two different sizes were predicted for clones containing the novel Sca I site (lanes 1, 2, 3, and 5). Some clones did not contain an insert that contained a sequence from the HIV-1 LTR and were not characterized further (lanes 7 and 8). (C) Sequence analysis of LM-PCR clones obtained from infected resting CD4 + T cells.

    Journal: Journal of Virology

    Article Title: Molecular Characterization of Preintegration Latency in Human Immunodeficiency Virus Type 1 Infection

    doi: 10.1128/JVI.76.17.8518-8531.2002

    Figure Lengend Snippet: Characterization of complete reverse transcripts cloned from resting T lymphocytes. (A) LM-PCR assay for complete reverse transcripts. The ligation of an asymmetric linker containing one-half of a Sca I site onto the terminus of a full-length HIV-1 genome creates a novel Sca I site, allowing rapid screening of cloned LM-PCR products. (B) Restriction analysis of cloned LM-PCR products obtained from resting CD4 + T cells. Highly purified resting CD4 + T cells were infected with HIV-1 IIIB. Three days after infection, cells were lysed and DNA was isolated for LM-PCR. Cloned LM-PCR products were restricted with Sca I and analyzed by Southern blotting using an LTR-specific probe. The analysis allowed the identification of blunt linear molecules with either the predicted U3 end (lanes 1, 2, 3, and 5) or clones bearing alterations that result in a change in the sequence of the most-terminal U3 nucleotides (lanes 4 and 6). Because the insert was cloned in either a forward or reverse orientation, inserts of two different sizes were predicted for clones containing the novel Sca I site (lanes 1, 2, 3, and 5). Some clones did not contain an insert that contained a sequence from the HIV-1 LTR and were not characterized further (lanes 7 and 8). (C) Sequence analysis of LM-PCR clones obtained from infected resting CD4 + T cells.

    Article Snippet: A mass of 200 ng of DNA was loaded into each PCR mixture, which contained 1 μM (each) primer, 200 μM deoxynucleoside triphosphates, 1.5 mM MgCl2 , and either 2.5 U of Expand High Fidelity Taq polymerase (Boehringer Mannheim) or 2.5 U of Platinum Taq (Life Technologies) in 1× proprietary buffer.

    Techniques: Clone Assay, Polymerase Chain Reaction, Ligation, Purification, Infection, Isolation, Southern Blot, Sequencing

    RT-PCR analysis of ERα and β transcripts in the heart chambers of newborn (4-day-old) and adult (55-day-old) female rats. Total RNA (1 μg) was reverse-transcribed, and cDNA was exponentially amplified in the presence of gene-specific primers. ( A ) A PhosphorImager scan after separation of the radioactive PCR product by electrophoresis in 1.5% agarose. Representative bands obtained from three RNA samples of the right atrium (RA) and left ventricle (LV) are shown. ( B ) Bar graphs depict ERα mRNA ( Left ) and ERβ mRNA ( Right ) levels verified by 18S RNA product used as internal standard. The results were calculated as a ratio of the signal given by radioactive bands of ERα mRNA or ERβ mRNA to the corresponding 18S band. RNA samples from the whole heart of newborn (1-day-old) rats were included in each experiment and used as a standard (100%) for calibration of ERα mRNA or ERβ mRNA levels. Values are means ± SEM for the right atrium and left ventricle, as well as for the left atrium (LA) and right ventricle (RV). n = 6; §, P

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

    Article Title: Estrogen receptors activate atrial natriuretic peptide in the rat heart

    doi: 10.1073/pnas.201394198

    Figure Lengend Snippet: RT-PCR analysis of ERα and β transcripts in the heart chambers of newborn (4-day-old) and adult (55-day-old) female rats. Total RNA (1 μg) was reverse-transcribed, and cDNA was exponentially amplified in the presence of gene-specific primers. ( A ) A PhosphorImager scan after separation of the radioactive PCR product by electrophoresis in 1.5% agarose. Representative bands obtained from three RNA samples of the right atrium (RA) and left ventricle (LV) are shown. ( B ) Bar graphs depict ERα mRNA ( Left ) and ERβ mRNA ( Right ) levels verified by 18S RNA product used as internal standard. The results were calculated as a ratio of the signal given by radioactive bands of ERα mRNA or ERβ mRNA to the corresponding 18S band. RNA samples from the whole heart of newborn (1-day-old) rats were included in each experiment and used as a standard (100%) for calibration of ERα mRNA or ERβ mRNA levels. Values are means ± SEM for the right atrium and left ventricle, as well as for the left atrium (LA) and right ventricle (RV). n = 6; §, P

    Article Snippet: Ten microliters of first-strand cDNA was added to a PCR mixture and amplified for 25–33 cycles by incubation at 95°C for 1 min, 57o C for 1 min, and 72°C for 1.5 min, with a final incubation at 72°C for 3 min, all in a Robocycler gradient 40 thermocycler (Stratagene).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Polymerase Chain Reaction, Electrophoresis

    ( A ) were subjected to SDS/PAGE, immunoblotted with an anti-ERα mAb, and visualized by the chemiluminescence technique. ( B and C ) RT-PCR analysis of ER transcripts and ANP in the left atrium ( B ) and right atrium ( C ) of sham-operated (sham), ovariectomized (ovx), and ovariectomized rats supplemented with 17-β-estradiol (ovxE). Total RNA (1 μg) was reverse-transcribed and cDNA was exponentially amplified in the presence of gene-specific primers. A PhosphorImager scan is presented after separation of the radioactive PCR product by electrophoresis in 1.5% agarose. Representative bands obtained in three RNA representative samples from sham, ovx, and ovxE rats are presented. The results were calculated as the ratio of the signal given by radioactive bands of ERα mRNA or ERβ mRNA to the corresponding 18S band. Values are means ± SEM adjusted to the values (in percentage) of sham-operated rats (100%). n = 6; *, P

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

    Article Title: Estrogen receptors activate atrial natriuretic peptide in the rat heart

    doi: 10.1073/pnas.201394198

    Figure Lengend Snippet: ( A ) were subjected to SDS/PAGE, immunoblotted with an anti-ERα mAb, and visualized by the chemiluminescence technique. ( B and C ) RT-PCR analysis of ER transcripts and ANP in the left atrium ( B ) and right atrium ( C ) of sham-operated (sham), ovariectomized (ovx), and ovariectomized rats supplemented with 17-β-estradiol (ovxE). Total RNA (1 μg) was reverse-transcribed and cDNA was exponentially amplified in the presence of gene-specific primers. A PhosphorImager scan is presented after separation of the radioactive PCR product by electrophoresis in 1.5% agarose. Representative bands obtained in three RNA representative samples from sham, ovx, and ovxE rats are presented. The results were calculated as the ratio of the signal given by radioactive bands of ERα mRNA or ERβ mRNA to the corresponding 18S band. Values are means ± SEM adjusted to the values (in percentage) of sham-operated rats (100%). n = 6; *, P

    Article Snippet: Ten microliters of first-strand cDNA was added to a PCR mixture and amplified for 25–33 cycles by incubation at 95°C for 1 min, 57o C for 1 min, and 72°C for 1.5 min, with a final incubation at 72°C for 3 min, all in a Robocycler gradient 40 thermocycler (Stratagene).

    Techniques: SDS Page, Reverse Transcription Polymerase Chain Reaction, Aqueous Normal-phase Chromatography, Amplification, Polymerase Chain Reaction, Electrophoresis

    Expression of survivin mRNA as assessed by RT-PCR in normal tissues, and oral cancer cell lines and primary oral cancer tissues . (A) Expression of survivin mRNA in oral cancer cell lines and primary oral cancer tissues from two patients. G3PDH expression was detected as an internal control. (B) Expression of survivin mRNA in normal adult tissues. 293T cells transfected with myc-tagged survivin cDNA (293T-survivin) was used as a positive control for survivin expression. G3PDH expression was detected as an internal control.

    Journal: Journal of Translational Medicine

    Article Title: Comparative study on the immunogenicity between an HLA-A24-restricted cytotoxic T-cell epitope derived from survivin and that from its splice variant survivin-2B in oral cancer patients

    doi: 10.1186/1479-5876-7-1

    Figure Lengend Snippet: Expression of survivin mRNA as assessed by RT-PCR in normal tissues, and oral cancer cell lines and primary oral cancer tissues . (A) Expression of survivin mRNA in oral cancer cell lines and primary oral cancer tissues from two patients. G3PDH expression was detected as an internal control. (B) Expression of survivin mRNA in normal adult tissues. 293T cells transfected with myc-tagged survivin cDNA (293T-survivin) was used as a positive control for survivin expression. G3PDH expression was detected as an internal control.

    Article Snippet: PCR amplification was performed in 50 ml of PCR mixture containing 1 mL of the cDNA mixture, KOD Plus DNA polymerase (Toyobo, Osaka, Japan), and 50 pmol of primers.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Positive Control

    Methylation-specific PCR results of some randomly selected sample patients with schizophrenia and control groups, analyzed with 2% agarose gel electrophoresis followed by ethidium bromide staining. A: Methylation pattern of some patients in CpG island-1 from BDNF gene. B: Methylation pattern of some controls in CpG island-1 from BDNF gene. C: Methylation pattern of some patients in CpG island-2 from BDNF gene. D: Methylation pattern of some controls in CpG island-2 from BDNF gene. PC: Positive control; M: methylated; U: unmethylated; 100M : 100 bp DNA ladder.

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: DNA Methylation of BDNF Gene in Schizophrenia

    doi: 10.12659/MSM.895896

    Figure Lengend Snippet: Methylation-specific PCR results of some randomly selected sample patients with schizophrenia and control groups, analyzed with 2% agarose gel electrophoresis followed by ethidium bromide staining. A: Methylation pattern of some patients in CpG island-1 from BDNF gene. B: Methylation pattern of some controls in CpG island-1 from BDNF gene. C: Methylation pattern of some patients in CpG island-2 from BDNF gene. D: Methylation pattern of some controls in CpG island-2 from BDNF gene. PC: Positive control; M: methylated; U: unmethylated; 100M : 100 bp DNA ladder.

    Article Snippet: PCR mixture contained 10 ng of modified DNA, 2.0 mmol each of dATP, dGTP, dCTP, and dTTP, 1.0 pmol each of primer, 2.0 mmol MgCl2, 1× reaction buffer, and 1.0 U Taq polymerase.

    Techniques: Methylation, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining, Positive Control

    Effects of HPV16 E1 mutants on HPV18 replication. (A) Schematic representation of HPV16 E1 mutants. F, FLAG-tag; ND, N-terminal domain; DBD, DNA-binding domain; OD, oligomerization domain; HD, helicase domain. (B) C33A cells were transfected with 1 ng of the HPV18 gDNA together with the indicated amounts (ng) of the expression plasmids. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with Dpn I. The Dpn I-resistant HPV18 gDNA was quantified by real-time PCR and normalized to the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication with the HPV18 E1/E2 alone. Each bar represents the average of two independent experiments with the standard error of mean.

    Journal: Virology Journal

    Article Title: Replication interference between human papillomavirus types 16 and 18 mediated by heterologous E1 helicases

    doi: 10.1186/1743-422X-11-11

    Figure Lengend Snippet: Effects of HPV16 E1 mutants on HPV18 replication. (A) Schematic representation of HPV16 E1 mutants. F, FLAG-tag; ND, N-terminal domain; DBD, DNA-binding domain; OD, oligomerization domain; HD, helicase domain. (B) C33A cells were transfected with 1 ng of the HPV18 gDNA together with the indicated amounts (ng) of the expression plasmids. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with Dpn I. The Dpn I-resistant HPV18 gDNA was quantified by real-time PCR and normalized to the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication with the HPV18 E1/E2 alone. Each bar represents the average of two independent experiments with the standard error of mean.

    Article Snippet: Total 20 μl of a PCR reaction mixture containing 2 μl of the Dpn I-digested sample, 10 μl of SYBR Green Realtime PCR Master Mix (TOYOBO CO., LTD, Osaka, Japan), and 0.4 μM of each primer was subjected to real-time PCR analysis using the LightCycler 480 (Roche Diagnostics).

    Techniques: FLAG-tag, Binding Assay, Transfection, Expressing, Molecular Weight, Isolation, Real-time Polymerase Chain Reaction, Luciferase

    Replication of HPV16 or HPV18 genomes supported by homologous or heterologous E1/E2s. C33A cells were transfected with 1 ng of the HPV16 (A) or HPV18 (B) gDNAs together with the indicated amounts (ng) of the expression plasmids for E1/E2s. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with Dpn I. The Dpn I-resistant HPV gDNA was quantified by real-time PCR and normalized to the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication when cells were transfected 20 ng of each pF16E1 and pF16E2 (A) or 10 ng of each pF18E1 and pF18E2 (B) . Each bar represents the average of two independent experiments with the standard error of mean.

    Journal: Virology Journal

    Article Title: Replication interference between human papillomavirus types 16 and 18 mediated by heterologous E1 helicases

    doi: 10.1186/1743-422X-11-11

    Figure Lengend Snippet: Replication of HPV16 or HPV18 genomes supported by homologous or heterologous E1/E2s. C33A cells were transfected with 1 ng of the HPV16 (A) or HPV18 (B) gDNAs together with the indicated amounts (ng) of the expression plasmids for E1/E2s. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with Dpn I. The Dpn I-resistant HPV gDNA was quantified by real-time PCR and normalized to the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication when cells were transfected 20 ng of each pF16E1 and pF16E2 (A) or 10 ng of each pF18E1 and pF18E2 (B) . Each bar represents the average of two independent experiments with the standard error of mean.

    Article Snippet: Total 20 μl of a PCR reaction mixture containing 2 μl of the Dpn I-digested sample, 10 μl of SYBR Green Realtime PCR Master Mix (TOYOBO CO., LTD, Osaka, Japan), and 0.4 μM of each primer was subjected to real-time PCR analysis using the LightCycler 480 (Roche Diagnostics).

    Techniques: Transfection, Expressing, Molecular Weight, Isolation, Real-time Polymerase Chain Reaction, Luciferase

    Simultaneous replication of HPV16/18 genomes in the presence of E1/E2s of both types. C33A cells were transfected with a mixture of HPV16 and HPV18 gDNAs (1 ng each) together with the indicated amounts (ng) of the expression plasmids. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with Dpn I. The Dpn I-resistant HPV16 (upper panel) and HPV18 (lower panel) gDNAs were quantified by real-time PCR and normalized to the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication with the homologous E1/E2 alone. Each bar represents the average of three independent experiments with the standard deviation.

    Journal: Virology Journal

    Article Title: Replication interference between human papillomavirus types 16 and 18 mediated by heterologous E1 helicases

    doi: 10.1186/1743-422X-11-11

    Figure Lengend Snippet: Simultaneous replication of HPV16/18 genomes in the presence of E1/E2s of both types. C33A cells were transfected with a mixture of HPV16 and HPV18 gDNAs (1 ng each) together with the indicated amounts (ng) of the expression plasmids. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with Dpn I. The Dpn I-resistant HPV16 (upper panel) and HPV18 (lower panel) gDNAs were quantified by real-time PCR and normalized to the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication with the homologous E1/E2 alone. Each bar represents the average of three independent experiments with the standard deviation.

    Article Snippet: Total 20 μl of a PCR reaction mixture containing 2 μl of the Dpn I-digested sample, 10 μl of SYBR Green Realtime PCR Master Mix (TOYOBO CO., LTD, Osaka, Japan), and 0.4 μM of each primer was subjected to real-time PCR analysis using the LightCycler 480 (Roche Diagnostics).

    Techniques: Transfection, Expressing, Molecular Weight, Isolation, Real-time Polymerase Chain Reaction, Luciferase, Standard Deviation

    Effects of heterologous E1 or E2 on HPV16/18 replication. C33A cells were transfected with 1 ng of the HPV16 (A) or HPV18 (B) gDNAs together with the indicated amounts (ng) of the expression plasmids for E1/E2s. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with Dpn I. The Dpn I-resistant HPV gDNA was quantified by real-time PCR and normalized to that of the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication with the homologous E1/E2 alone. Each bar represents the average of two independent experiments with the standard error of mean.

    Journal: Virology Journal

    Article Title: Replication interference between human papillomavirus types 16 and 18 mediated by heterologous E1 helicases

    doi: 10.1186/1743-422X-11-11

    Figure Lengend Snippet: Effects of heterologous E1 or E2 on HPV16/18 replication. C33A cells were transfected with 1 ng of the HPV16 (A) or HPV18 (B) gDNAs together with the indicated amounts (ng) of the expression plasmids for E1/E2s. Three days after transfection, low molecular weight DNA was isolated by the Hirt procedure and digested with Dpn I. The Dpn I-resistant HPV gDNA was quantified by real-time PCR and normalized to that of the luciferase gene. The level of the replication was presented as the relative amount of the Dpn I-resistant DNA compared to that obtained by the replication with the homologous E1/E2 alone. Each bar represents the average of two independent experiments with the standard error of mean.

    Article Snippet: Total 20 μl of a PCR reaction mixture containing 2 μl of the Dpn I-digested sample, 10 μl of SYBR Green Realtime PCR Master Mix (TOYOBO CO., LTD, Osaka, Japan), and 0.4 μM of each primer was subjected to real-time PCR analysis using the LightCycler 480 (Roche Diagnostics).

    Techniques: Transfection, Expressing, Molecular Weight, Isolation, Real-time Polymerase Chain Reaction, Luciferase

    Standard curve generated from quantification cycle numbers of a 10-fold dilution series of an amplicon product of 86 bp cloned into a pCR 2.1 plasmid vector in DNase- and RNase-free water (≤5.00 to 5.00 × 10 5 copies/reaction). qPCR was

    Journal: Applied and Environmental Microbiology

    Article Title: Enumeration of Salmonellae in Table Eggs, Pasteurized Egg Products, and Egg-Containing Dishes by Using Quantitative Real-Time PCR

    doi: 10.1128/AEM.03360-13

    Figure Lengend Snippet: Standard curve generated from quantification cycle numbers of a 10-fold dilution series of an amplicon product of 86 bp cloned into a pCR 2.1 plasmid vector in DNase- and RNase-free water (≤5.00 to 5.00 × 10 5 copies/reaction). qPCR was

    Article Snippet: The cycling conditions were 95°C for 20 s and then 45 cycles of 95°C for 3 s and 65°C for 30 s. All DNA samples were tested in triplicate in a Peltier-based real-time PCR instrument (Applied Biosystems 7500 Fast real-time PCR system). (ii) A 25-μl PCR mixture contained 12.5 μl of Fast qPCR Mastermix Plus–No ROX (Eurogentec, Seraing, Belgium); 1 μl of IAC, kindly provided by the Bundesinstitut für Risikobewertung (BfR) (Federal Institute for Risk Assessment) ( ); 0.4 μM each primer (ttr4 and ttr-6; Eurofins MWG Operon, Germany); 0.24 μM the LNA target probe; 0.24 μM the IAC probe (Yakima Yellow-CACACGGCGACGCGAACGCTTT-BHQ1) (Eurogentec) ( ); and 5 μl of template DNA.

    Techniques: Generated, Amplification, Clone Assay, Polymerase Chain Reaction, Plasmid Preparation, Real-time Polymerase Chain Reaction

    Survival of S . Enteritidis strain MB2509 in tiramisu. DNA was extracted by using the DNeasy Blood and Tissue kit, and qPCR was performed with the Applied Biosystems 7500 Fast real-time PCR system. Asterisks indicate significantly different numbers of

    Journal: Applied and Environmental Microbiology

    Article Title: Enumeration of Salmonellae in Table Eggs, Pasteurized Egg Products, and Egg-Containing Dishes by Using Quantitative Real-Time PCR

    doi: 10.1128/AEM.03360-13

    Figure Lengend Snippet: Survival of S . Enteritidis strain MB2509 in tiramisu. DNA was extracted by using the DNeasy Blood and Tissue kit, and qPCR was performed with the Applied Biosystems 7500 Fast real-time PCR system. Asterisks indicate significantly different numbers of

    Article Snippet: The cycling conditions were 95°C for 20 s and then 45 cycles of 95°C for 3 s and 65°C for 30 s. All DNA samples were tested in triplicate in a Peltier-based real-time PCR instrument (Applied Biosystems 7500 Fast real-time PCR system). (ii) A 25-μl PCR mixture contained 12.5 μl of Fast qPCR Mastermix Plus–No ROX (Eurogentec, Seraing, Belgium); 1 μl of IAC, kindly provided by the Bundesinstitut für Risikobewertung (BfR) (Federal Institute for Risk Assessment) ( ); 0.4 μM each primer (ttr4 and ttr-6; Eurofins MWG Operon, Germany); 0.24 μM the LNA target probe; 0.24 μM the IAC probe (Yakima Yellow-CACACGGCGACGCGAACGCTTT-BHQ1) (Eurogentec) ( ); and 5 μl of template DNA.

    Techniques: Real-time Polymerase Chain Reaction

    Genomic organization of MsHid . A. Schematic diagram of exon-intron structure. The horizontal boxes indicate exons (1 st exon, 657 bp; 2 nd exon, 145 bp; 3 rd exon, 205 bp; 4 th exon 249 bp). The locations of three introns are indicated by vertical lines that are accompanied with partial sequences (consensus 5′ and 3′ splicing junctions are indicated in reds). Arrows indicate PCR primers. A Hin P1 I site was used for inverse PCR. B. An agarose gel electrophoresis confirming introns. Primer sets are as follows: lane 1 and 2, 5′ORF-R2; lane 3, F3-Int1R; lane 4 and 5, RTF1-R1; lane 6, RTF1 and Int2R; lane 7 and 8, RTF2 and RTR2; lane 9, RTF2 and Int3R. Note that there is no PCR fragment in lane 1 and 7 due to large size of the 1 st and the 3 rd introns, respectively. However, PCR fragments from genomic DNA using the intron-derived primers match expected sizes (lane 3, 6, 9). (abbr.: G, genomic DNA template; C, cDNA template). C. Comparison of gene structure between DmHid (top) and MsHid (bottom). Exons are indicated by boxes, introns by lines. Numbers indicate nucleotide lengths of introns (asterisks for approximate values).

    Journal: Gene

    Article Title: Cloning and Functional Characterizations of an Apoptogenic Hid Gene in the Scuttle Fly, Megaselia scalaris (Diptera; Phoridae)

    doi: 10.1016/j.gene.2016.11.043

    Figure Lengend Snippet: Genomic organization of MsHid . A. Schematic diagram of exon-intron structure. The horizontal boxes indicate exons (1 st exon, 657 bp; 2 nd exon, 145 bp; 3 rd exon, 205 bp; 4 th exon 249 bp). The locations of three introns are indicated by vertical lines that are accompanied with partial sequences (consensus 5′ and 3′ splicing junctions are indicated in reds). Arrows indicate PCR primers. A Hin P1 I site was used for inverse PCR. B. An agarose gel electrophoresis confirming introns. Primer sets are as follows: lane 1 and 2, 5′ORF-R2; lane 3, F3-Int1R; lane 4 and 5, RTF1-R1; lane 6, RTF1 and Int2R; lane 7 and 8, RTF2 and RTR2; lane 9, RTF2 and Int3R. Note that there is no PCR fragment in lane 1 and 7 due to large size of the 1 st and the 3 rd introns, respectively. However, PCR fragments from genomic DNA using the intron-derived primers match expected sizes (lane 3, 6, 9). (abbr.: G, genomic DNA template; C, cDNA template). C. Comparison of gene structure between DmHid (top) and MsHid (bottom). Exons are indicated by boxes, introns by lines. Numbers indicate nucleotide lengths of introns (asterisks for approximate values).

    Article Snippet: Briefly, 50 μL of PCR mixture contained 0.5 μg of genomic DNA, 2 μM forward and reverse primer (5 μL each), 2.5 μL of 10 mM dNTPs, 5 μL of 10X tuning buffer, and 0.4 μL of polymerase (5 PRIME).

    Techniques: Polymerase Chain Reaction, Inverse PCR, Agarose Gel Electrophoresis, Derivative Assay

    PCR assay for detection of bla VEB gene with product size: 600 bp. Lanes 3–17: Amplified products, Lanes 2 and 19: Positive control, Lanes 1 and 20: DNA ladder 100 bp, Lane 18: Negative control

    Journal: GMS Hygiene and Infection Control

    Article Title: Prevalence of extended-spectrum beta-lactamase genes in Acinetobacter baumannii strains isolated from nosocomial infections in Tehran, Iran

    doi: 10.3205/dgkh000318

    Figure Lengend Snippet: PCR assay for detection of bla VEB gene with product size: 600 bp. Lanes 3–17: Amplified products, Lanes 2 and 19: Positive control, Lanes 1 and 20: DNA ladder 100 bp, Lane 18: Negative control

    Article Snippet: The PCR mixture contained the DNA template, Forward/Reverse primers, and master mix (Bioneer Co., Korea, Cat. number K-2016).

    Techniques: Polymerase Chain Reaction, Amplification, Positive Control, Negative Control

    PCR assay for detection of bla TEM gene with product size: 700 bp. Lanes 2–36: Amplified products, Lane 37: Positive control, Lanes 1 and 40: DNA ladder 100 bp, Lane 38–40: Negative control

    Journal: GMS Hygiene and Infection Control

    Article Title: Prevalence of extended-spectrum beta-lactamase genes in Acinetobacter baumannii strains isolated from nosocomial infections in Tehran, Iran

    doi: 10.3205/dgkh000318

    Figure Lengend Snippet: PCR assay for detection of bla TEM gene with product size: 700 bp. Lanes 2–36: Amplified products, Lane 37: Positive control, Lanes 1 and 40: DNA ladder 100 bp, Lane 38–40: Negative control

    Article Snippet: The PCR mixture contained the DNA template, Forward/Reverse primers, and master mix (Bioneer Co., Korea, Cat. number K-2016).

    Techniques: Polymerase Chain Reaction, Transmission Electron Microscopy, Amplification, Positive Control, Negative Control

    PCR assay for detection of bla SHV gene with product size: 700 bp. Lanes 3–18: Amplified products, Lane 2: Positive control, Lanes 1 and 20: DNA ladder 100 bp, Lane 19: Negative control

    Journal: GMS Hygiene and Infection Control

    Article Title: Prevalence of extended-spectrum beta-lactamase genes in Acinetobacter baumannii strains isolated from nosocomial infections in Tehran, Iran

    doi: 10.3205/dgkh000318

    Figure Lengend Snippet: PCR assay for detection of bla SHV gene with product size: 700 bp. Lanes 3–18: Amplified products, Lane 2: Positive control, Lanes 1 and 20: DNA ladder 100 bp, Lane 19: Negative control

    Article Snippet: The PCR mixture contained the DNA template, Forward/Reverse primers, and master mix (Bioneer Co., Korea, Cat. number K-2016).

    Techniques: Polymerase Chain Reaction, Amplification, Positive Control, Negative Control