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  • 99
    Thermo Fisher pcr mixture
    Transcriptional analysis of the ces gene cluster. (A) Bars indicate overlapping primer pairs used for transcriptional analysis of the ces operon (listed in Table S1 in the supplemental material). Bent arrows indicate promoters as determined by primer extension. (B) <t>RT-PCR</t> showed the presence of consecutive transcripts between cesP and cesD (lane 3 to 9) but no transcripts between cesH and cesP (lane 2). There were no transcripts from a forward primer in cesD and reverse primer downstream of cesD after the predicted hairpin termination structure (lane 10). Negative controls (RNA; see lane H-) and positive controls (DNA; see lane H+). M, marker ladder mixture (Fermentas).
    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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    Bio-Rad pcr mixture
    Inhibition of vaccinia virus transcription by Tkip and SOCS1-KIR. BSC-40 cells mock treated or treated with lipo-Tkip, lipo-SOCS1-KIR, or alanine substitution-containing control peptides for 1 h were infected with vaccinia virus at an MOI of 5 for 1 h. The cells were washed and incubated in growth medium containing the same concentration of peptides for 18 h. RNA was extracted and used for <t>cDNA</t> synthesis, followed by quantitative <t>PCR.</t> The expression of early (D12L) (a), intermediate (AIL) (b), and late (A7L) (c) genes was compared with endogenous actin gene expression.
    Pcr Mixture, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 1531 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad pcr reaction mixture
    HPLC separation and electropherogram of the <t>PCR</t> products from the ErbB1 oncogene and the interferon gamma (INFγ) gene for the detection of the ErbB1 gene amplification. A) Chromatograms showing the results for normal <t>DNA</t> (control), DNA from breast tumors (test) and BT-474 breast carcinoma cell line (high copy control); B) Lanes 1, 2-3, 4 and 5 are respectively blank, controls, high copy control and DNA from primary breast carcinoma. The PCR products of INFγ and ErbB1 genes are very close together
    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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    Meridian Life Science pcr mixture
    ( a ) Transcriptional analysis of the A. pleuropneumoniae NGT locus. Lane 1: <t>cDNA</t> as template; lane 2: RNA as template; lane 3: A. pleuropneumoniae HS143 genomic DNA positive control; lane 4: negative <t>PCR</t> control (no template). ( b ) Transcriptional analysis of the region upstream of ngt . Lane 1: cDNA as template; lane 2: genomic DNA positive control; lane 3: PCR control (RNA as template).
    Pcr Mixture, supplied by Meridian Life Science, used in various techniques. Bioz Stars score: 94/100, based on 772 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boehringer Mannheim pcr mixture
    (A) Recombination between heterologous RNA during replication results in four proviral forms, the two parental forms, NL4-3Δ pol and NL4-3Δ env , and two recombinant forms, wild-type NL4-3 (NL4-3WT) and NL4-3Δ pol/Δenv . Arrows denote oligonucleotides used for <t>PCR</t> amplification <t>(Rec1,</t> Rec2, DIS1, and DIS2) and hybridization (Std*, Pol *, and ENV *) and the direction of sequence complementarity. Oligonucleotides marked with asterisks are probes for detecting PCR products. Hatch marks denote unrepresented sequences in HIV-1. Long terminal repeat (LTR) and structural genes are shown. (B) Provirus resulting from a single-cycle infection of SupT1 cells. Wild-type NL4-3 DIS (CG) and virus generated by cotransfection of NL4-3Δ pol DIS (CG) and NL4-3Δ env DIS (CG) were used to infect SupT1 cells. Lysates of infected cells were amplified with primers Rec1 and Rec2, and proviral sequences were identified by hybridization with Std*, Pol *, and ENV * probes. Molecular weights of the PCR products confirm expected recombination products. (C) DNA recombination during virus production. Hirt supernatants from 293 cells transfected with no DNA, pNL4-3Δ env DIS (CG), pNL4-3Δ env DIS (CG) and NL4-3Δ pol DIS (CG), NL4-3Δ pol DIS (CG), or NL4-3 DIS (CG) were amplified with primers Rec1 and Rec2, and proviral sequences were identified by hybridization with the Std* probe. Lysate of SupT1 cells infected (Inf) with virus generated by cotransfection of NL4-3Δ pol DIS (CG) and NL4-3Δ env DIS (CG) was amplified as a control. WT, wild type.
    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
    Pre-RC proteins are associated with region B in the origin of bidirectional replication (OBR). ( A ) Scheme of the origin of replication in murine rDNA. The transcription start site is marked as +1. OBR designates the origin of bidirectional replication mapped by nascent strand determination analysis ( 32 ). <t>DNA</t> fragments used in ChIP and EMSA assays are denoted according to the nucleotide position at the 5′ and at the 3′ ends relative to the transcription start site. The 119 bp fragment used in EMSA experiments is marked by arrows. The conspicuous 9 bp repetitive sequence elements are underlined. ( B ) Cross-linked chromatin was immunoprecipitated with antibodies against ORC2 (αORC2) and preimmune serum (control). Isolated DNA was analyzed with primer pairs flanking regions A, B, C and D, respectively. <t>PCR</t> products of the corresponding chromatin samples before immunoprecipitation are shown (input).
    Pcr Mixture, supplied by Stratagene, used in various techniques. Bioz Stars score: 93/100, 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 pcr reaction mixture
    Identification of putative miRNAs targeting the 3′UTR of hTERT . a Expression of hTERT mRNA in different cell lines as quantified by real-time <t>RT-PCR.</t> The expression of hTERT mRNA in WA01 human ES cells is set as 1. b Telomerase activity in different cell lines as quantified by real-time <t>TRAP.</t> The telomerase activity in WA01 human ES cells is set as 1. c – e Relative Renilla/Firefly luciferase luminescence in different cells transiently transfected with psiCHECK2-5′UTR, psiCHECK2-3′UTR or psiCHECK2-5′ + 3′UTR vector comparing to the same cells transiently transfected with psiCHECK2 empty vector. f Z score distribution of the log 2 Renilla/Firefly luciferase luminescence ratio from three independent screens using a miRNA inhibitor library in HeLa cells stably expressing psiCHECK2-5′ + 3′UTR reporter. Candidates with Z score > 2.7 were chosen for further analysis. The p value from one-sided test is 0.0034. g – i Relative Renilla/Firefly luciferase luminescence, hTERT mRNA expression and telomerase activity in HeLa cells transiently transfected with each of the eight candidate miRNA inhibitors. The miRNA candidates with reported differential expression in stem cells and terminal differentiated cells are highlighted in red
    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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    Thermo Fisher taqman universal pcr master mixture
    PRMT5 overexpression in lung cancer is evident at mRNA and protein levels. (a) There is a 6.13-fold increase in PRMT5 mRNA levels in lung tumors (LT), 8 cases, over matched nonneoplastic pulmonary parenchyma (L), as evident by <t>TaqMan</t> real time polymerase chain reaction <t>(PCR).</t> (b) PRMT5 and its symmetric methylation mark H4R3 are detected in lung carcinoma cell lines (NCI-A549, NCI-H520) but not in the nonneoplastic human pulmonary alveolar (HPAEpiC) and bronchial epithelial cell lines (HBEpiC); cellular localization experiments highlight nuclear and cytoplasmic fractions of PRMT5; Western immunoblot (b, c) ; immunohistochemistry (d) , NCI-H69, original magnification ×600.
    Taqman Universal Pcr Master Mixture, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 352 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co pcr mixture
    Analysis of <t>PCR</t> products by agarose gel electrophoresis. M, Marker. (A) B19 <t>DNA</t> was amplified using nested-PCR as described in Materials and methods. Lanes 1, 2, 3, 6, 7 and 10, six seropositive samples; lanes 4, 5, 8 and 9, four seronegative samples;
    Pcr Mixture, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 92/100, based on 349 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    ATUM pcr mixture
    Diagram representing <t>PCR-amplified</t> DNA containing the mononucleotide <t>SSR</t> locus 1 (underlined). Restriction enzyme cut sites are presented as boxed arrowheads along with the corresponding masses of each fragment.
    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
    The expression profile of all three Kindlin paralogs was quantified by real-time <t>RT-PCR</t> using <t>SYBR</t> Green. Data are presented as the relative expression of Kindlins (fold change) normalized by a housekeeping gene, β-actin. Note: The error bars indicate the standard deviation.
    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
    Amount of Synechococcus <t>DNA</t> as a function of density as determined by CsCl centrifugation and quantitative <t>PCR,</t> using the Synechococcus rbcL gene as a proxy in field samples. The vertical lines correspond to the vertical lines in Fig. and
    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
    Amount of Synechococcus <t>DNA</t> as a function of density as determined by CsCl centrifugation and quantitative <t>PCR,</t> using the Synechococcus rbcL gene as a proxy in field samples. The vertical lines correspond to the vertical lines in Fig. and
    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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    Meridian Life Science pcr reaction mixture
    <t>16S</t> rRNA polymerase chain reaction product of Streptococcus agalactiae (Lane 1) amplified at 1500 bp with standard molecular weight marker (M 1 kb; M 100 bp) and negative control (NC).
    Pcr Reaction Mixture, supplied by Meridian Life Science, used in various techniques. Bioz Stars score: 92/100, based on 195 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: 93/100, based on 169 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bioneer Corporation pcr mixture
    Polymerase Chain Reaction Amplification of the adeC Gene of the A. baumannii Isolates Lane M, 100 bp <t>DNA</t> size marker; Lane P, A. baumannii ATCC19606 positive control; Lane N, negative control; Lane 1 and 2, adeC gene positive isolate.
    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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    Thermo Fisher taqman pcr mixture
    HIF-1α activates the induction of BCL-9 expression by hypoxia Knockdown of endogenous HIF-1α but not HIF-2α largely abolishes the induction of BCL-9 expression by hypoxia in HCT116 cells. Cells with knockdown of endogenous HIF-1α, HIF-2α by siRNA oligos or transfected with control siRNA were treated with hypoxia for 36 h. Two different siRNA oligos against HIF-1α and HIF-2α, respectively, were used, and similar results were obtained. ( A ) The mRNA expression levels of BCL-9 (left panel) and VEGF (right panel) were determined by <t>Taqman</t> real-time <t>PCR</t> and normalized with actin. ( B ) The BCL-9 protein levels were determined by Western-blot assays. ( C ) The knockdown of HIF-2α (left panel) and HIF-1α (right panel) in cells was conformed at the mRNA level by Taqman real-time PCR and normalized with actin. Data are presented as mean ± SD ( n = 3). * p
    Taqman Pcr Mixture, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 191 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: 93/100, based on 189 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TransGen biotech co 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 TransGen biotech co, used in various techniques. Bioz Stars score: 93/100, based on 178 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega 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 Promega, used in various techniques. Bioz Stars score: 94/100, based on 3256 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman fast universal pcr master 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).
    Taqman Fast Universal Pcr Master Mixture, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 211 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Transcriptional analysis of the ces gene cluster. (A) Bars indicate overlapping primer pairs used for transcriptional analysis of the ces operon (listed in Table S1 in the supplemental material). Bent arrows indicate promoters as determined by primer extension. (B) RT-PCR showed the presence of consecutive transcripts between cesP and cesD (lane 3 to 9) but no transcripts between cesH and cesP (lane 2). There were no transcripts from a forward primer in cesD and reverse primer downstream of cesD after the predicted hairpin termination structure (lane 10). Negative controls (RNA; see lane H-) and positive controls (DNA; see lane H+). M, marker ladder mixture (Fermentas).

    Journal: Applied and Environmental Microbiology

    Article Title: Identification of the Main Promoter Directing Cereulide Biosynthesis in Emetic Bacillus cereus and Its Application for Real-Time Monitoring of ces Gene Expression in Foods ▿ Gene Expression in Foods ▿ †

    doi: 10.1128/AEM.02317-09

    Figure Lengend Snippet: Transcriptional analysis of the ces gene cluster. (A) Bars indicate overlapping primer pairs used for transcriptional analysis of the ces operon (listed in Table S1 in the supplemental material). Bent arrows indicate promoters as determined by primer extension. (B) RT-PCR showed the presence of consecutive transcripts between cesP and cesD (lane 3 to 9) but no transcripts between cesH and cesP (lane 2). There were no transcripts from a forward primer in cesD and reverse primer downstream of cesD after the predicted hairpin termination structure (lane 10). Negative controls (RNA; see lane H-) and positive controls (DNA; see lane H+). M, marker ladder mixture (Fermentas).

    Article Snippet: The 50-μl PCR mixture (10 ng DNA, 0.5 μM each primer, 1.5 mM MgCl2 , 0.4 mM each deoxynucleoside triphosphate [dNTP], 1.25 U ThermoStart Taq polymerase [all from ABgene]) was activated (95°C for 15 min), followed by 30 amplification cycles (95°C for 30 s, 60°C for 45 s, and 72°C for 1 min) and an elongation step (72°C for 5 min).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Marker

    Inhibition of vaccinia virus transcription by Tkip and SOCS1-KIR. BSC-40 cells mock treated or treated with lipo-Tkip, lipo-SOCS1-KIR, or alanine substitution-containing control peptides for 1 h were infected with vaccinia virus at an MOI of 5 for 1 h. The cells were washed and incubated in growth medium containing the same concentration of peptides for 18 h. RNA was extracted and used for cDNA synthesis, followed by quantitative PCR. The expression of early (D12L) (a), intermediate (AIL) (b), and late (A7L) (c) genes was compared with endogenous actin gene expression.

    Journal: Journal of Virology

    Article Title: SOCS-1 Mimetics Protect Mice against Lethal Poxvirus Infection: Identification of a Novel Endogenous Antiviral System ▿

    doi: 10.1128/JVI.01138-08

    Figure Lengend Snippet: Inhibition of vaccinia virus transcription by Tkip and SOCS1-KIR. BSC-40 cells mock treated or treated with lipo-Tkip, lipo-SOCS1-KIR, or alanine substitution-containing control peptides for 1 h were infected with vaccinia virus at an MOI of 5 for 1 h. The cells were washed and incubated in growth medium containing the same concentration of peptides for 18 h. RNA was extracted and used for cDNA synthesis, followed by quantitative PCR. The expression of early (D12L) (a), intermediate (AIL) (b), and late (A7L) (c) genes was compared with endogenous actin gene expression.

    Article Snippet: The cDNA was diluted 1:100, and 1 μl of the diluted cDNA was taken in a 50-μl reaction mixture with primers and a PCR mixture containing SYBR green (Bio-Rad, Hercules, CA) and used for PCR in a Mini Opticon thermal cycler (Bio-Rad, Hercules, CA).

    Techniques: Inhibition, Infection, Incubation, Concentration Assay, Real-time Polymerase Chain Reaction, Expressing

    HPLC separation and electropherogram of the PCR products from the ErbB1 oncogene and the interferon gamma (INFγ) gene for the detection of the ErbB1 gene amplification. A) Chromatograms showing the results for normal DNA (control), DNA from breast tumors (test) and BT-474 breast carcinoma cell line (high copy control); B) Lanes 1, 2-3, 4 and 5 are respectively blank, controls, high copy control and DNA from primary breast carcinoma. The PCR products of INFγ and ErbB1 genes are very close together

    Journal: Avicenna Journal of Medical Biotechnology

    Article Title: Quantitative Analysis of ErbB1 and ErbB2 Genes Amplification by a High Performance Liquid Chromatography

    doi:

    Figure Lengend Snippet: HPLC separation and electropherogram of the PCR products from the ErbB1 oncogene and the interferon gamma (INFγ) gene for the detection of the ErbB1 gene amplification. A) Chromatograms showing the results for normal DNA (control), DNA from breast tumors (test) and BT-474 breast carcinoma cell line (high copy control); B) Lanes 1, 2-3, 4 and 5 are respectively blank, controls, high copy control and DNA from primary breast carcinoma. The PCR products of INFγ and ErbB1 genes are very close together

    Article Snippet: The PCR reaction mixture contained 3 µl of genomic DNA (100 ng , 5-fold diluted), 1 µl of 5 mmol/l solutions of each of the forward and reverse primers, and 12.5 µl of 2 × SYBR green DNA PCR Master Mix (BioRad, USA) in a total volume of 25 µl .

    Techniques: High Performance Liquid Chromatography, Polymerase Chain Reaction, Amplification

    HPLC separation and electropherogram of The PCR products from the ErbB2 oncogene and the interferon gamma gene (INFγ) for the detection of the ErbB2 gene amplification. A) Chromatograms showing the results for normal DNA (control), DNA from cancerous breast (test) and BT-474 breast carcinoma cell line (high copy control); B) Lane 1 is high copy control, lanes 2 and 3 are DNA from primary breast carcinoma and lane 4 is normal breast tissue

    Journal: Avicenna Journal of Medical Biotechnology

    Article Title: Quantitative Analysis of ErbB1 and ErbB2 Genes Amplification by a High Performance Liquid Chromatography

    doi:

    Figure Lengend Snippet: HPLC separation and electropherogram of The PCR products from the ErbB2 oncogene and the interferon gamma gene (INFγ) for the detection of the ErbB2 gene amplification. A) Chromatograms showing the results for normal DNA (control), DNA from cancerous breast (test) and BT-474 breast carcinoma cell line (high copy control); B) Lane 1 is high copy control, lanes 2 and 3 are DNA from primary breast carcinoma and lane 4 is normal breast tissue

    Article Snippet: The PCR reaction mixture contained 3 µl of genomic DNA (100 ng , 5-fold diluted), 1 µl of 5 mmol/l solutions of each of the forward and reverse primers, and 12.5 µl of 2 × SYBR green DNA PCR Master Mix (BioRad, USA) in a total volume of 25 µl .

    Techniques: High Performance Liquid Chromatography, Polymerase Chain Reaction, Amplification

    ( a ) Transcriptional analysis of the A. pleuropneumoniae NGT locus. Lane 1: cDNA as template; lane 2: RNA as template; lane 3: A. pleuropneumoniae HS143 genomic DNA positive control; lane 4: negative PCR control (no template). ( b ) Transcriptional analysis of the region upstream of ngt . Lane 1: cDNA as template; lane 2: genomic DNA positive control; lane 3: PCR control (RNA as template).

    Journal: Open Biology

    Article Title: The N-linking glycosylation system from Actinobacillus pleuropneumoniae is required for adhesion and has potential use in glycoengineering

    doi: 10.1098/rsob.160212

    Figure Lengend Snippet: ( a ) Transcriptional analysis of the A. pleuropneumoniae NGT locus. Lane 1: cDNA as template; lane 2: RNA as template; lane 3: A. pleuropneumoniae HS143 genomic DNA positive control; lane 4: negative PCR control (no template). ( b ) Transcriptional analysis of the region upstream of ngt . Lane 1: cDNA as template; lane 2: genomic DNA positive control; lane 3: PCR control (RNA as template).

    Article Snippet: For each sample, 2 µl of reverse transcribed cDNA was used as a template in a 25 µl total volume PCR mixture and amplified using MyTaq master mix (Bioline, UK) using the following cycling conditions: 94°C/30 s, followed by 35 cycles of 94°C/10 s, 53°C/10 s, 72°C/10 s and a final single 72°C/30 s cycle using the primers listed in electronic supplementary material, table S2.

    Techniques: Positive Control, Polymerase Chain Reaction

    (A) Recombination between heterologous RNA during replication results in four proviral forms, the two parental forms, NL4-3Δ pol and NL4-3Δ env , and two recombinant forms, wild-type NL4-3 (NL4-3WT) and NL4-3Δ pol/Δenv . Arrows denote oligonucleotides used for PCR amplification (Rec1, Rec2, DIS1, and DIS2) and hybridization (Std*, Pol *, and ENV *) and the direction of sequence complementarity. Oligonucleotides marked with asterisks are probes for detecting PCR products. Hatch marks denote unrepresented sequences in HIV-1. Long terminal repeat (LTR) and structural genes are shown. (B) Provirus resulting from a single-cycle infection of SupT1 cells. Wild-type NL4-3 DIS (CG) and virus generated by cotransfection of NL4-3Δ pol DIS (CG) and NL4-3Δ env DIS (CG) were used to infect SupT1 cells. Lysates of infected cells were amplified with primers Rec1 and Rec2, and proviral sequences were identified by hybridization with Std*, Pol *, and ENV * probes. Molecular weights of the PCR products confirm expected recombination products. (C) DNA recombination during virus production. Hirt supernatants from 293 cells transfected with no DNA, pNL4-3Δ env DIS (CG), pNL4-3Δ env DIS (CG) and NL4-3Δ pol DIS (CG), NL4-3Δ pol DIS (CG), or NL4-3 DIS (CG) were amplified with primers Rec1 and Rec2, and proviral sequences were identified by hybridization with the Std* probe. Lysate of SupT1 cells infected (Inf) with virus generated by cotransfection of NL4-3Δ pol DIS (CG) and NL4-3Δ env DIS (CG) was amplified as a control. WT, wild type.

    Journal: Journal of Virology

    Article Title: Infectious Molecular Clones with the Nonhomologous Dimer Initiation Sequences Found in Different Subtypes of Human Immunodeficiency Virus Type 1 Can Recombine and Initiate a Spreading Infection In Vitro

    doi:

    Figure Lengend Snippet: (A) Recombination between heterologous RNA during replication results in four proviral forms, the two parental forms, NL4-3Δ pol and NL4-3Δ env , and two recombinant forms, wild-type NL4-3 (NL4-3WT) and NL4-3Δ pol/Δenv . Arrows denote oligonucleotides used for PCR amplification (Rec1, Rec2, DIS1, and DIS2) and hybridization (Std*, Pol *, and ENV *) and the direction of sequence complementarity. Oligonucleotides marked with asterisks are probes for detecting PCR products. Hatch marks denote unrepresented sequences in HIV-1. Long terminal repeat (LTR) and structural genes are shown. (B) Provirus resulting from a single-cycle infection of SupT1 cells. Wild-type NL4-3 DIS (CG) and virus generated by cotransfection of NL4-3Δ pol DIS (CG) and NL4-3Δ env DIS (CG) were used to infect SupT1 cells. Lysates of infected cells were amplified with primers Rec1 and Rec2, and proviral sequences were identified by hybridization with Std*, Pol *, and ENV * probes. Molecular weights of the PCR products confirm expected recombination products. (C) DNA recombination during virus production. Hirt supernatants from 293 cells transfected with no DNA, pNL4-3Δ env DIS (CG), pNL4-3Δ env DIS (CG) and NL4-3Δ pol DIS (CG), NL4-3Δ pol DIS (CG), or NL4-3 DIS (CG) were amplified with primers Rec1 and Rec2, and proviral sequences were identified by hybridization with the Std* probe. Lysate of SupT1 cells infected (Inf) with virus generated by cotransfection of NL4-3Δ pol DIS (CG) and NL4-3Δ env DIS (CG) was amplified as a control. WT, wild type.

    Article Snippet: The PCR mixture contained 250 nM primers Rec1 and Rec2, 350 μM deoxynucleoside triphosphate, 1.75 mM MgCl2 , 50 mM Tris-HCl (pH 9.2), 16 mM (NH4 )2 SO4 , and 3.5 U of polymerase (Boehringer Mannheim).

    Techniques: Recombinant, Polymerase Chain Reaction, Amplification, Hybridization, Sequencing, Infection, Generated, Cotransfection, Transfection

    Pre-RC proteins are associated with region B in the origin of bidirectional replication (OBR). ( A ) Scheme of the origin of replication in murine rDNA. The transcription start site is marked as +1. OBR designates the origin of bidirectional replication mapped by nascent strand determination analysis ( 32 ). DNA fragments used in ChIP and EMSA assays are denoted according to the nucleotide position at the 5′ and at the 3′ ends relative to the transcription start site. The 119 bp fragment used in EMSA experiments is marked by arrows. The conspicuous 9 bp repetitive sequence elements are underlined. ( B ) Cross-linked chromatin was immunoprecipitated with antibodies against ORC2 (αORC2) and preimmune serum (control). Isolated DNA was analyzed with primer pairs flanking regions A, B, C and D, respectively. PCR products of the corresponding chromatin samples before immunoprecipitation are shown (input).

    Journal: Nucleic Acids Research

    Article Title: Site-specific interaction of the murine pre-replicative complex with origin DNA: assembly and disassembly during cell cycle transit and differentiation

    doi: 10.1093/nar/gkm555

    Figure Lengend Snippet: Pre-RC proteins are associated with region B in the origin of bidirectional replication (OBR). ( A ) Scheme of the origin of replication in murine rDNA. The transcription start site is marked as +1. OBR designates the origin of bidirectional replication mapped by nascent strand determination analysis ( 32 ). DNA fragments used in ChIP and EMSA assays are denoted according to the nucleotide position at the 5′ and at the 3′ ends relative to the transcription start site. The 119 bp fragment used in EMSA experiments is marked by arrows. The conspicuous 9 bp repetitive sequence elements are underlined. ( B ) Cross-linked chromatin was immunoprecipitated with antibodies against ORC2 (αORC2) and preimmune serum (control). Isolated DNA was analyzed with primer pairs flanking regions A, B, C and D, respectively. PCR products of the corresponding chromatin samples before immunoprecipitation are shown (input).

    Article Snippet: Site-directed mutagenesis The PCR mixture contained 20 ng template plasmid DNA (1545 bp rDNA fragment from −3476 to −1931 cloned into pUC18), 15 pmol of each oligonucleotide, 200 µM dNTPs, 2.5 U Pfu DNA polymerase (Stratagene) and buffer in a total volume of 50 µl.

    Techniques: Chromatin Immunoprecipitation, Sequencing, Immunoprecipitation, Isolation, Polymerase Chain Reaction

    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

    Identification of putative miRNAs targeting the 3′UTR of hTERT . a Expression of hTERT mRNA in different cell lines as quantified by real-time RT-PCR. The expression of hTERT mRNA in WA01 human ES cells is set as 1. b Telomerase activity in different cell lines as quantified by real-time TRAP. The telomerase activity in WA01 human ES cells is set as 1. c – e Relative Renilla/Firefly luciferase luminescence in different cells transiently transfected with psiCHECK2-5′UTR, psiCHECK2-3′UTR or psiCHECK2-5′ + 3′UTR vector comparing to the same cells transiently transfected with psiCHECK2 empty vector. f Z score distribution of the log 2 Renilla/Firefly luciferase luminescence ratio from three independent screens using a miRNA inhibitor library in HeLa cells stably expressing psiCHECK2-5′ + 3′UTR reporter. Candidates with Z score > 2.7 were chosen for further analysis. The p value from one-sided test is 0.0034. g – i Relative Renilla/Firefly luciferase luminescence, hTERT mRNA expression and telomerase activity in HeLa cells transiently transfected with each of the eight candidate miRNA inhibitors. The miRNA candidates with reported differential expression in stem cells and terminal differentiated cells are highlighted in red

    Journal: Nature Communications

    Article Title: HoxC5 and miR-615-3p target newly evolved genomic regions to repress hTERT and inhibit tumorigenesis

    doi: 10.1038/s41467-017-02601-1

    Figure Lengend Snippet: Identification of putative miRNAs targeting the 3′UTR of hTERT . a Expression of hTERT mRNA in different cell lines as quantified by real-time RT-PCR. The expression of hTERT mRNA in WA01 human ES cells is set as 1. b Telomerase activity in different cell lines as quantified by real-time TRAP. The telomerase activity in WA01 human ES cells is set as 1. c – e Relative Renilla/Firefly luciferase luminescence in different cells transiently transfected with psiCHECK2-5′UTR, psiCHECK2-3′UTR or psiCHECK2-5′ + 3′UTR vector comparing to the same cells transiently transfected with psiCHECK2 empty vector. f Z score distribution of the log 2 Renilla/Firefly luciferase luminescence ratio from three independent screens using a miRNA inhibitor library in HeLa cells stably expressing psiCHECK2-5′ + 3′UTR reporter. Candidates with Z score > 2.7 were chosen for further analysis. The p value from one-sided test is 0.0034. g – i Relative Renilla/Firefly luciferase luminescence, hTERT mRNA expression and telomerase activity in HeLa cells transiently transfected with each of the eight candidate miRNA inhibitors. The miRNA candidates with reported differential expression in stem cells and terminal differentiated cells are highlighted in red

    Article Snippet: The PCR reaction mixture contained 1X TRAP buffer, 4 ng/μl ACX primer 5′-GCGCGGCTTA CCCTTACCCTTACCCTAACC-3′, 15% glycerol, 1:2000 SYBR Green from Invitrogen, 0.08 U/μl Taq polymerase and was initiated at 94 °C for 2 min, followed by 45 cycles of 95 °C for 5 s, 50 °C for 6 s, and 72 °C for 10 s. The threshold cycle values (Ct) were determined from semi log amplification plots.

    Techniques: Expressing, Quantitative RT-PCR, Activity Assay, Luciferase, Transfection, Plasmid Preparation, Stable Transfection

    PRMT5 overexpression in lung cancer is evident at mRNA and protein levels. (a) There is a 6.13-fold increase in PRMT5 mRNA levels in lung tumors (LT), 8 cases, over matched nonneoplastic pulmonary parenchyma (L), as evident by TaqMan real time polymerase chain reaction (PCR). (b) PRMT5 and its symmetric methylation mark H4R3 are detected in lung carcinoma cell lines (NCI-A549, NCI-H520) but not in the nonneoplastic human pulmonary alveolar (HPAEpiC) and bronchial epithelial cell lines (HBEpiC); cellular localization experiments highlight nuclear and cytoplasmic fractions of PRMT5; Western immunoblot (b, c) ; immunohistochemistry (d) , NCI-H69, original magnification ×600.

    Journal: Diagnostic Pathology

    Article Title: Cellular localization of protein arginine methyltransferase-5 correlates with grade of lung tumors

    doi: 10.1186/1746-1596-8-201

    Figure Lengend Snippet: PRMT5 overexpression in lung cancer is evident at mRNA and protein levels. (a) There is a 6.13-fold increase in PRMT5 mRNA levels in lung tumors (LT), 8 cases, over matched nonneoplastic pulmonary parenchyma (L), as evident by TaqMan real time polymerase chain reaction (PCR). (b) PRMT5 and its symmetric methylation mark H4R3 are detected in lung carcinoma cell lines (NCI-A549, NCI-H520) but not in the nonneoplastic human pulmonary alveolar (HPAEpiC) and bronchial epithelial cell lines (HBEpiC); cellular localization experiments highlight nuclear and cytoplasmic fractions of PRMT5; Western immunoblot (b, c) ; immunohistochemistry (d) , NCI-H69, original magnification ×600.

    Article Snippet: The PCR amplification was conducted in 25 μl reaction using the TaqMan Universal PCR Master Mixture (Applied Biosystems, Foster City, CA) according to the protocol supplied by the manufacturer.

    Techniques: Over Expression, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Methylation, Western Blot, Immunohistochemistry

    Analysis of PCR products by agarose gel electrophoresis. M, Marker. (A) B19 DNA was amplified using nested-PCR as described in Materials and methods. Lanes 1, 2, 3, 6, 7 and 10, six seropositive samples; lanes 4, 5, 8 and 9, four seronegative samples;

    Journal: Oncology Letters

    Article Title: High prevelance of human parvovirus infection in patients with malignant tumors

    doi: 10.3892/ol.2012.548

    Figure Lengend Snippet: Analysis of PCR products by agarose gel electrophoresis. M, Marker. (A) B19 DNA was amplified using nested-PCR as described in Materials and methods. Lanes 1, 2, 3, 6, 7 and 10, six seropositive samples; lanes 4, 5, 8 and 9, four seronegative samples;

    Article Snippet: The extracted DNA (2.5 μl) was added to the PCR mixture containing 2.5 μl of 10X reaction buffer (Tiangen Biotech, Beijing Co. Ltd, China), 200 μM of each dATP, dCTP, dGTP and dTTP, 12.5 pmol of each primer and 1.25 units of Taq polymerase (Tiangen Biotech).

    Techniques: Polymerase Chain Reaction, Agarose Gel Electrophoresis, Marker, Amplification, Nested PCR

    Diagram representing PCR-amplified DNA containing the mononucleotide SSR locus 1 (underlined). Restriction enzyme cut sites are presented as boxed arrowheads along with the corresponding masses of each fragment.

    Journal: Applied and Environmental Microbiology

    Article Title: Improved Short-Sequence-Repeat Genotyping of Mycobacterium avium subsp. paratuberculosis by Using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

    doi: 10.1128/AEM.03212-13

    Figure Lengend Snippet: Diagram representing PCR-amplified DNA containing the mononucleotide SSR locus 1 (underlined). Restriction enzyme cut sites are presented as boxed arrowheads along with the corresponding masses of each fragment.

    Article Snippet: The SSR locus 1 was amplified in a PCR mixture containing 2.5 units of Herculase enhanced DNA polymerase, 200 μM (each) deoxynucleoside triphosphate (dNTP), 1.5 pmol forward primer, 3.0 pmol reverse primer, 5 μl 10× buffer, and 2 μl template DNA (20 to 40 ng) in a total volume of 50 μl.

    Techniques: Polymerase Chain Reaction, Amplification

    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

    The expression profile of all three Kindlin paralogs was quantified by real-time RT-PCR using SYBR Green. Data are presented as the relative expression of Kindlins (fold change) normalized by a housekeeping gene, β-actin. Note: The error bars indicate the standard deviation.

    Journal: Evolutionary Bioinformatics Online

    Article Title: Phylogenetic Analysis of Kindlins Suggests Subfunctionalization of an Ancestral Unduplicated Kindlin into Three Paralogs in Vertebrates

    doi: 10.4137/EBO.S6179

    Figure Lengend Snippet: The expression profile of all three Kindlin paralogs was quantified by real-time RT-PCR using SYBR Green. Data are presented as the relative expression of Kindlins (fold change) normalized by a housekeeping gene, β-actin. Note: The error bars indicate the standard deviation.

    Article Snippet: Triplicate samples of each PCR mixture, each containing 4.7 μl of POWER SYBR Green PCR master mixture (Applied Biosystems), 0.3 μl of a 10 pmol/μl of primer mixture, 0.3 μl of cDNA, and water to a total volume of 10 μl were transferred into a 96-well plate on an ABI 7500 Fast Real Time PCR System (Applied Biosystems).

    Techniques: Expressing, Quantitative RT-PCR, SYBR Green Assay, Standard Deviation

    Amount of Synechococcus DNA as a function of density as determined by CsCl centrifugation and quantitative PCR, using the Synechococcus rbcL gene as a proxy in field samples. The vertical lines correspond to the vertical lines in Fig. and

    Journal: Applied and Environmental Microbiology

    Article Title: Use of Inorganic and Organic Nitrogen by Synechococcus spp. and Diatoms on the West Florida Shelf as Measured Using Stable Isotope Probing ▿ spp. and Diatoms on the West Florida Shelf as Measured Using Stable Isotope Probing ▿ †

    doi: 10.1128/AEM.01002-09

    Figure Lengend Snippet: Amount of Synechococcus DNA as a function of density as determined by CsCl centrifugation and quantitative PCR, using the Synechococcus rbcL gene as a proxy in field samples. The vertical lines correspond to the vertical lines in Fig. and

    Article Snippet: Two microliters of DNA from each fraction was added to 28 μl of PCR master mixture prepared using 2× PCR TaqMan master mixture (PE Applied Biosystems, Foster City, CA) containing each primer at a concentration of 1 μM, 2 mM MgCl2 , and 100 nM probe.

    Techniques: Centrifugation, Real-time Polymerase Chain Reaction

    16S rRNA polymerase chain reaction product of Streptococcus agalactiae (Lane 1) amplified at 1500 bp with standard molecular weight marker (M 1 kb; M 100 bp) and negative control (NC).

    Journal: Veterinary World

    Article Title: Molecular identification and histopathological study of natural Streptococcus agalactiae infection in hybrid tilapia (Oreochromis niloticus)

    doi: 10.14202/vetworld.2017.101-111

    Figure Lengend Snippet: 16S rRNA polymerase chain reaction product of Streptococcus agalactiae (Lane 1) amplified at 1500 bp with standard molecular weight marker (M 1 kb; M 100 bp) and negative control (NC).

    Article Snippet: PCR The PCR reaction mixture of 16s rRNA was done in 25 µl total reaction using ×2 MyTaq mix (Bioline, UK) with 10 µM of each primer.

    Techniques: Polymerase Chain Reaction, Amplification, Molecular Weight, Marker, Negative Control

    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

    Polymerase Chain Reaction Amplification of the adeC Gene of the A. baumannii Isolates Lane M, 100 bp DNA size marker; Lane P, A. baumannii ATCC19606 positive control; Lane N, negative control; Lane 1 and 2, adeC gene positive isolate.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Efflux Pump Inhibitor Phenylalanine-Arginine Β-Naphthylamide Effect on the Minimum Inhibitory Concentration of Imipenem in Acinetobacter baumannii Strains Isolated From Hospitalized Patients in Shahid Motahari Burn Hospital, Tehran, Iran

    doi: 10.5812/jjm.19048

    Figure Lengend Snippet: Polymerase Chain Reaction Amplification of the adeC Gene of the A. baumannii Isolates Lane M, 100 bp DNA size marker; Lane P, A. baumannii ATCC19606 positive control; Lane N, negative control; Lane 1 and 2, adeC gene positive isolate.

    Article Snippet: Briefly, the 25 µL PCR mixture contained 2.5 µL of bacterial DNA, 10 pM of each primer, 1.5 mM of MgCl2 , 250 µM of each dNTP, 10 Mm of Tris-HCL (pH: 9.0), 30 Mm of KCL, and 1 U of Taq DNA polymerase (Bioneer Company, Korea, Cat. No. K-2012).

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

    Polymerase Chain Reaction Amplification of the adeB Gene of the A. baumannii Isolates Lane M, 100 bp DNA size marker; Lane P, A. baumannii ATCC19606 positive control; Lane N, negative control; Lane 1 - 14, adeB gene positive isolate.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Efflux Pump Inhibitor Phenylalanine-Arginine Β-Naphthylamide Effect on the Minimum Inhibitory Concentration of Imipenem in Acinetobacter baumannii Strains Isolated From Hospitalized Patients in Shahid Motahari Burn Hospital, Tehran, Iran

    doi: 10.5812/jjm.19048

    Figure Lengend Snippet: Polymerase Chain Reaction Amplification of the adeB Gene of the A. baumannii Isolates Lane M, 100 bp DNA size marker; Lane P, A. baumannii ATCC19606 positive control; Lane N, negative control; Lane 1 - 14, adeB gene positive isolate.

    Article Snippet: Briefly, the 25 µL PCR mixture contained 2.5 µL of bacterial DNA, 10 pM of each primer, 1.5 mM of MgCl2 , 250 µM of each dNTP, 10 Mm of Tris-HCL (pH: 9.0), 30 Mm of KCL, and 1 U of Taq DNA polymerase (Bioneer Company, Korea, Cat. No. K-2012).

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

    Polymerase Chain Reaction Amplification of the adeA Gene of the A. baumannii Isolates Lane M, 100 bp DNA size marker; Lane P, A. baumannii ATCC19606 positive control; Lane N, negative control; Lane 1 - 7, adeA gene positive isolate.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Efflux Pump Inhibitor Phenylalanine-Arginine Β-Naphthylamide Effect on the Minimum Inhibitory Concentration of Imipenem in Acinetobacter baumannii Strains Isolated From Hospitalized Patients in Shahid Motahari Burn Hospital, Tehran, Iran

    doi: 10.5812/jjm.19048

    Figure Lengend Snippet: Polymerase Chain Reaction Amplification of the adeA Gene of the A. baumannii Isolates Lane M, 100 bp DNA size marker; Lane P, A. baumannii ATCC19606 positive control; Lane N, negative control; Lane 1 - 7, adeA gene positive isolate.

    Article Snippet: Briefly, the 25 µL PCR mixture contained 2.5 µL of bacterial DNA, 10 pM of each primer, 1.5 mM of MgCl2 , 250 µM of each dNTP, 10 Mm of Tris-HCL (pH: 9.0), 30 Mm of KCL, and 1 U of Taq DNA polymerase (Bioneer Company, Korea, Cat. No. K-2012).

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

    HIF-1α activates the induction of BCL-9 expression by hypoxia Knockdown of endogenous HIF-1α but not HIF-2α largely abolishes the induction of BCL-9 expression by hypoxia in HCT116 cells. Cells with knockdown of endogenous HIF-1α, HIF-2α by siRNA oligos or transfected with control siRNA were treated with hypoxia for 36 h. Two different siRNA oligos against HIF-1α and HIF-2α, respectively, were used, and similar results were obtained. ( A ) The mRNA expression levels of BCL-9 (left panel) and VEGF (right panel) were determined by Taqman real-time PCR and normalized with actin. ( B ) The BCL-9 protein levels were determined by Western-blot assays. ( C ) The knockdown of HIF-2α (left panel) and HIF-1α (right panel) in cells was conformed at the mRNA level by Taqman real-time PCR and normalized with actin. Data are presented as mean ± SD ( n = 3). * p

    Journal: Oncotarget

    Article Title: HIF-1α activates hypoxia-induced BCL-9 expression in human colorectal cancer cells

    doi: 10.18632/oncotarget.8834

    Figure Lengend Snippet: HIF-1α activates the induction of BCL-9 expression by hypoxia Knockdown of endogenous HIF-1α but not HIF-2α largely abolishes the induction of BCL-9 expression by hypoxia in HCT116 cells. Cells with knockdown of endogenous HIF-1α, HIF-2α by siRNA oligos or transfected with control siRNA were treated with hypoxia for 36 h. Two different siRNA oligos against HIF-1α and HIF-2α, respectively, were used, and similar results were obtained. ( A ) The mRNA expression levels of BCL-9 (left panel) and VEGF (right panel) were determined by Taqman real-time PCR and normalized with actin. ( B ) The BCL-9 protein levels were determined by Western-blot assays. ( C ) The knockdown of HIF-2α (left panel) and HIF-1α (right panel) in cells was conformed at the mRNA level by Taqman real-time PCR and normalized with actin. Data are presented as mean ± SD ( n = 3). * p

    Article Snippet: Real-time PCR was done in triplicate with TaqMan PCR mixture (Applied Biosystems).

    Techniques: Expressing, Transfection, Real-time Polymerase Chain Reaction, Western Blot

    Hypoxia induces BCL-9 expression levels in human colorectal cancer cell lines Human colorectal cancer cell lines SW480 and HCT116 cells were cultured under the hypoxic condition for the indicated time periods. ( A ) The mRNA expression levels of BCL-9 in these cells were determined by Taqman real-time PCR and normalized with actin. ( B ) The mRNA expression levels of VEGF in these cells were determined as a positive control. ( C ) The BCL-9 protein levels were determined by Western-blot assays. Data are presented as mean ± SD ( n = 3). * p

    Journal: Oncotarget

    Article Title: HIF-1α activates hypoxia-induced BCL-9 expression in human colorectal cancer cells

    doi: 10.18632/oncotarget.8834

    Figure Lengend Snippet: Hypoxia induces BCL-9 expression levels in human colorectal cancer cell lines Human colorectal cancer cell lines SW480 and HCT116 cells were cultured under the hypoxic condition for the indicated time periods. ( A ) The mRNA expression levels of BCL-9 in these cells were determined by Taqman real-time PCR and normalized with actin. ( B ) The mRNA expression levels of VEGF in these cells were determined as a positive control. ( C ) The BCL-9 protein levels were determined by Western-blot assays. Data are presented as mean ± SD ( n = 3). * p

    Article Snippet: Real-time PCR was done in triplicate with TaqMan PCR mixture (Applied Biosystems).

    Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Positive Control, Western Blot

    HIF-1α transcriptionally stimulates BCL-9 expression ( A ) Ectopic HIF-1α expression increases BCL-9 and VEGF mRNA expression levels in SW480 and HCT116 cells. The mRNA expression levels of BCL-9 and VEGF were determined by Taqman real-time PCR and normalized with actin. ( B ) Ectopic HIF-2α expression increases VEGF mRNA expression levels but has no effect on mRNA BCL-9 expression levels in SW480 and HCT116 cells. ( C ) Ectopic HIF-1α expression increases BCL-9 protein levels in SW480 and HCT116 cells as determined by Western-blot assays. ( D ) and ( E ) HIF-1α binds to HRE-B and HRE-C sites in the BCL-9 promoter in HCT116 cells transfected with HIF-1α expression plasmids as determined by ChIP assays. The amount of DNA fragments pulled-down was determined by real-time PCR (D) or conventional PCR (E). The HRE site in the VEGF promoter serves as a positive control. ( F ) HIF-1α activates luciferase activity of reporter vectors containing HRE-B or HRE-C sites in the BCL-9 promoter in HCT116 cells transfected with HIF-1α expression plasmids. Luciferase reporter vectors containing the HRE site in the VEGF promoter was included as a positive control. Data are presented as mean ± SD ( n = 3). * p

    Journal: Oncotarget

    Article Title: HIF-1α activates hypoxia-induced BCL-9 expression in human colorectal cancer cells

    doi: 10.18632/oncotarget.8834

    Figure Lengend Snippet: HIF-1α transcriptionally stimulates BCL-9 expression ( A ) Ectopic HIF-1α expression increases BCL-9 and VEGF mRNA expression levels in SW480 and HCT116 cells. The mRNA expression levels of BCL-9 and VEGF were determined by Taqman real-time PCR and normalized with actin. ( B ) Ectopic HIF-2α expression increases VEGF mRNA expression levels but has no effect on mRNA BCL-9 expression levels in SW480 and HCT116 cells. ( C ) Ectopic HIF-1α expression increases BCL-9 protein levels in SW480 and HCT116 cells as determined by Western-blot assays. ( D ) and ( E ) HIF-1α binds to HRE-B and HRE-C sites in the BCL-9 promoter in HCT116 cells transfected with HIF-1α expression plasmids as determined by ChIP assays. The amount of DNA fragments pulled-down was determined by real-time PCR (D) or conventional PCR (E). The HRE site in the VEGF promoter serves as a positive control. ( F ) HIF-1α activates luciferase activity of reporter vectors containing HRE-B or HRE-C sites in the BCL-9 promoter in HCT116 cells transfected with HIF-1α expression plasmids. Luciferase reporter vectors containing the HRE site in the VEGF promoter was included as a positive control. Data are presented as mean ± SD ( n = 3). * p

    Article Snippet: Real-time PCR was done in triplicate with TaqMan PCR mixture (Applied Biosystems).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Transfection, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Positive Control, Luciferase, Activity Assay

    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