polymerase chain reaction pcr fragment Search Results


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  • 99
    New England Biolabs polymerase chain reaction restriction fragment length polymorphism pcr rflp
    Polymerase Chain Reaction Restriction Fragment Length Polymorphism Pcr Rflp, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore polymerase chain reaction pcr fragment
    Polymerase Chain Reaction Pcr Fragment, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GeneDx Inc polymerase chain reaction amplified fragments
    Polymerase Chain Reaction Amplified Fragments, supplied by GeneDx Inc, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche pcr fragments
    The example of <t>HRM</t> analysis in patients 385 and 419 (exon 39). 1 Detection of mutations by HRM. The green curve in the diagram corresponds to the <t>PCR</t> fragment carrying the nonsense mutation c.11172G > A (p.Trp3724X) in patient 385. The red curve refers to the PCR fragment carrying the likely pathogenic substitution c.11248C > G (p.Arg3750Gly) in patient 419. The rest of fragments with blue curves are wild type samples (patients without sequence changes). 2, 3 Confirmation of mutations by direct sequencing in patient 385 (number 2) and patient 419 (number 3). Both sequences are in reverse direction. The letter A corresponds to the mutated sequence; B is the wild type.
    Pcr Fragments, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 1197 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa polymerase chain reaction pcr fragments
    Ectopically expressed intronless <t>TSG101</t> gene (cDNA) is spliced and generates equivalent product as the endogenous aberrant mRNA in cancer cells. ( A ) The structure of the TSG101 mRNA. Red line and triangles indicate the postulated mRNA re-splicing via activated alternative 5′ and 3′ splice sites. Blue triangle indicates the PTC (1112–1114) generated by the postulated splicing. Black flags indicate the open reading frame (ORF; 127–1296). The positions of the <t>PCR</t> primers (P1–P4, P7–P10) are shown. ( B ) Using normal cells (HMEC) and cancer cells (MCF-7), endogenous TSG101 mRNAs were analyzed by RT-PCR (RT) and the genomic DNA was analyzed by PCR (Ge) with primer sets (P1–P4, P7–P10) annealed to the indicated exons. The black and red arrowheads denote the full-length and major aberrant mRNAs, respectively ( Figure 1 ). ( C ) The structures of three reporter TSG101-EGFP plasmids (see (A) for the red and blue triangles). The postulated splicing-dependent EGFP expression of these plasmids is indicated with (−) and (+). The positions of the PCR primer sets (P1–P5, P3–P6) are shown. ( D ) Cancer cells (MCF-7) were transfected with the indicated TSG101-EGFP reporter plasmids and the EGFP expression was analyzed by fluorescence microscopy. ( E ) These ectopically expressed TSG101-EGFP transcripts were analyzed by RT-PCR. The black and red arrowheads indicate the unspliced and spliced products via the alternative splice sites, respectively. Two minor spliced products (indicated with * and **) were generated from different alternative 3′ splice sites, i.e. Δ190–776 (587-nt deletion) and Δ190–1236 (1047-nt deletion), respectively.
    Polymerase Chain Reaction Pcr Fragments, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad polymerase chain reaction pcr fragment
    Real-time <t>RT-PCR</t> analyses of <t>tHsc70</t> expression in the liver (a), skeletal muscle (b), lung (c), and heart (d) of P . sinensis after heat stress for 1, 2, and 4 h with 1-h adaptive recovery at 25 °C (Groups V, VI, and VII) or without adaptation
    Polymerase Chain Reaction Pcr Fragment, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 88/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare pcr fragments
    Relative expression profiles of down- (A) or up-regulated (B) monolignol genes in F . × ananassa cv. Elsanta . Total RNA was isolated from a single untreated fruit (wild type; WT), control fruits (pBI-intron), down-regulated fruits (pBI- FaCCRi , pBI- FaCADi , and pBI- FaPODi ) (A) , and up-regulated fruits (pBI- <t>FaCCR</t> , pBI- FaCAD , and pBI- FaPOD ) (B) . Expression levels of all samples were monitored by <t>qRT-PCR</t> with specific primers for target genes ( FaCCR . FaCAD , and FaPOD ) and an interspac er gene. The latter was used as an internal control for normalization. For each box-plot graph, one of the WT group was used as the reference (set to one) and each group contained five biological replicates. The Wilcoxon-Mann-Whitney U -test was used for a non-parametric comparison of two groups from pBI-intron and fruits infiltrated with different constructs. Values indicate statistically decreased and increased levels ( P
    Pcr Fragments, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 93/100, based on 2054 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Axygen polymerase chain reaction fragments
    Relative expression profiles of down- (A) or up-regulated (B) monolignol genes in F . × ananassa cv. Elsanta . Total RNA was isolated from a single untreated fruit (wild type; WT), control fruits (pBI-intron), down-regulated fruits (pBI- FaCCRi , pBI- FaCADi , and pBI- FaPODi ) (A) , and up-regulated fruits (pBI- <t>FaCCR</t> , pBI- FaCAD , and pBI- FaPOD ) (B) . Expression levels of all samples were monitored by <t>qRT-PCR</t> with specific primers for target genes ( FaCCR . FaCAD , and FaPOD ) and an interspac er gene. The latter was used as an internal control for normalization. For each box-plot graph, one of the WT group was used as the reference (set to one) and each group contained five biological replicates. The Wilcoxon-Mann-Whitney U -test was used for a non-parametric comparison of two groups from pBI-intron and fruits infiltrated with different constructs. Values indicate statistically decreased and increased levels ( P
    Polymerase Chain Reaction Fragments, supplied by Axygen, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa pcr fragment
    <t>MEF2C</t> expression and PSL sensitivity of primary leukemic blast cells in four T-ALL patients (two ETP-ALL and two non-ETP ALL). (a) MEF2C expression levels determined by real-time <t>quantitative-PCR.</t> GAPDH was used as an internal control. (b) Viability of cells was counted after treatment with PSL (200 μM) and/or ABT-737 (10 nM). Annexin (-)/PI(-) cells were defined as viable cells.
    Pcr Fragment, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 2963 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad polymerase chain reaction fragments
    <t>MEF2C</t> expression and PSL sensitivity of primary leukemic blast cells in four T-ALL patients (two ETP-ALL and two non-ETP ALL). (a) MEF2C expression levels determined by real-time <t>quantitative-PCR.</t> GAPDH was used as an internal control. (b) Viability of cells was counted after treatment with PSL (200 μM) and/or ABT-737 (10 nM). Annexin (-)/PI(-) cells were defined as viable cells.
    Polymerase Chain Reaction Fragments, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Illumina Inc pcr fragments
    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative <t>PCR</t> (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from <t>MiSeq</t> analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P
    Pcr Fragments, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 92/100, based on 486 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Eurofins polymerase chain reaction pcr fragments
    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative <t>PCR</t> (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from <t>MiSeq</t> analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P
    Polymerase Chain Reaction Pcr Fragments, supplied by Eurofins, used in various techniques. Bioz Stars score: 89/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene polymerase chain reaction pcr fragment
    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative <t>PCR</t> (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from <t>MiSeq</t> analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P
    Polymerase Chain Reaction Pcr Fragment, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche polymerase chain reaction pcr fragment
    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative <t>PCR</t> (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from <t>MiSeq</t> analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P
    Polymerase Chain Reaction Pcr Fragment, supplied by Roche, used in various techniques. Bioz Stars score: 88/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene polymerase chain reaction pcr fragments
    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative <t>PCR</t> (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from <t>MiSeq</t> analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P
    Polymerase Chain Reaction Pcr Fragments, supplied by Stratagene, used in various techniques. Bioz Stars score: 89/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MJ Research polymerase chain reaction restriction fragment length polymorphism pcr rflp
    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative <t>PCR</t> (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from <t>MiSeq</t> analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P
    Polymerase Chain Reaction Restriction Fragment Length Polymorphism Pcr Rflp, supplied by MJ Research, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche polymerase chain reaction pcr amplified fragments
    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative <t>PCR</t> (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from <t>MiSeq</t> analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P
    Polymerase Chain Reaction Pcr Amplified Fragments, supplied by Roche, used in various techniques. Bioz Stars score: 85/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    4Gene polymerase chain reaction fragment length polymorphism
    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative <t>PCR</t> (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from <t>MiSeq</t> analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P
    Polymerase Chain Reaction Fragment Length Polymorphism, supplied by 4Gene, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher polymerase chain reaction restriction fragment length polymorphism pcr rflp assay
    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative <t>PCR</t> (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from <t>MiSeq</t> analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P
    Polymerase Chain Reaction Restriction Fragment Length Polymorphism Pcr Rflp Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore polymerase chain reaction pcr
    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative <t>PCR</t> (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from <t>MiSeq</t> analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P
    Polymerase Chain Reaction Pcr, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 363 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bio-Rad polymerase chain reaction restriction fragment length polymorphism
    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative <t>PCR</t> (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from <t>MiSeq</t> analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P
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    The example of HRM analysis in patients 385 and 419 (exon 39). 1 Detection of mutations by HRM. The green curve in the diagram corresponds to the PCR fragment carrying the nonsense mutation c.11172G > A (p.Trp3724X) in patient 385. The red curve refers to the PCR fragment carrying the likely pathogenic substitution c.11248C > G (p.Arg3750Gly) in patient 419. The rest of fragments with blue curves are wild type samples (patients without sequence changes). 2, 3 Confirmation of mutations by direct sequencing in patient 385 (number 2) and patient 419 (number 3). Both sequences are in reverse direction. The letter A corresponds to the mutated sequence; B is the wild type.

    Journal: BMC Medical Genetics

    Article Title: Novel mutations of PKD genes in the Czech population with autosomal dominant polycystic kidney disease

    doi: 10.1186/1471-2350-15-41

    Figure Lengend Snippet: The example of HRM analysis in patients 385 and 419 (exon 39). 1 Detection of mutations by HRM. The green curve in the diagram corresponds to the PCR fragment carrying the nonsense mutation c.11172G > A (p.Trp3724X) in patient 385. The red curve refers to the PCR fragment carrying the likely pathogenic substitution c.11248C > G (p.Arg3750Gly) in patient 419. The rest of fragments with blue curves are wild type samples (patients without sequence changes). 2, 3 Confirmation of mutations by direct sequencing in patient 385 (number 2) and patient 419 (number 3). Both sequences are in reverse direction. The letter A corresponds to the mutated sequence; B is the wild type.

    Article Snippet: PCR fragments were produced and analyzed by HRM using the LightCycler® 480 (Roche Applied Science).

    Techniques: Polymerase Chain Reaction, Mutagenesis, Sequencing

    Ectopically expressed intronless TSG101 gene (cDNA) is spliced and generates equivalent product as the endogenous aberrant mRNA in cancer cells. ( A ) The structure of the TSG101 mRNA. Red line and triangles indicate the postulated mRNA re-splicing via activated alternative 5′ and 3′ splice sites. Blue triangle indicates the PTC (1112–1114) generated by the postulated splicing. Black flags indicate the open reading frame (ORF; 127–1296). The positions of the PCR primers (P1–P4, P7–P10) are shown. ( B ) Using normal cells (HMEC) and cancer cells (MCF-7), endogenous TSG101 mRNAs were analyzed by RT-PCR (RT) and the genomic DNA was analyzed by PCR (Ge) with primer sets (P1–P4, P7–P10) annealed to the indicated exons. The black and red arrowheads denote the full-length and major aberrant mRNAs, respectively ( Figure 1 ). ( C ) The structures of three reporter TSG101-EGFP plasmids (see (A) for the red and blue triangles). The postulated splicing-dependent EGFP expression of these plasmids is indicated with (−) and (+). The positions of the PCR primer sets (P1–P5, P3–P6) are shown. ( D ) Cancer cells (MCF-7) were transfected with the indicated TSG101-EGFP reporter plasmids and the EGFP expression was analyzed by fluorescence microscopy. ( E ) These ectopically expressed TSG101-EGFP transcripts were analyzed by RT-PCR. The black and red arrowheads indicate the unspliced and spliced products via the alternative splice sites, respectively. Two minor spliced products (indicated with * and **) were generated from different alternative 3′ splice sites, i.e. Δ190–776 (587-nt deletion) and Δ190–1236 (1047-nt deletion), respectively.

    Journal: Nucleic Acids Research

    Article Title: Re-splicing of mature mRNA in cancer cells promotes activation of distant weak alternative splice sites

    doi: 10.1093/nar/gks520

    Figure Lengend Snippet: Ectopically expressed intronless TSG101 gene (cDNA) is spliced and generates equivalent product as the endogenous aberrant mRNA in cancer cells. ( A ) The structure of the TSG101 mRNA. Red line and triangles indicate the postulated mRNA re-splicing via activated alternative 5′ and 3′ splice sites. Blue triangle indicates the PTC (1112–1114) generated by the postulated splicing. Black flags indicate the open reading frame (ORF; 127–1296). The positions of the PCR primers (P1–P4, P7–P10) are shown. ( B ) Using normal cells (HMEC) and cancer cells (MCF-7), endogenous TSG101 mRNAs were analyzed by RT-PCR (RT) and the genomic DNA was analyzed by PCR (Ge) with primer sets (P1–P4, P7–P10) annealed to the indicated exons. The black and red arrowheads denote the full-length and major aberrant mRNAs, respectively ( Figure 1 ). ( C ) The structures of three reporter TSG101-EGFP plasmids (see (A) for the red and blue triangles). The postulated splicing-dependent EGFP expression of these plasmids is indicated with (−) and (+). The positions of the PCR primer sets (P1–P5, P3–P6) are shown. ( D ) Cancer cells (MCF-7) were transfected with the indicated TSG101-EGFP reporter plasmids and the EGFP expression was analyzed by fluorescence microscopy. ( E ) These ectopically expressed TSG101-EGFP transcripts were analyzed by RT-PCR. The black and red arrowheads indicate the unspliced and spliced products via the alternative splice sites, respectively. Two minor spliced products (indicated with * and **) were generated from different alternative 3′ splice sites, i.e. Δ190–776 (587-nt deletion) and Δ190–1236 (1047-nt deletion), respectively.

    Article Snippet: Construction of plasmids The expression plasmids for the TSG101-EGFP fusion proteins (TSG101-EGFP, TSG101-EGFP[+] and TSG101-EGFP[−]; C) were constructed by subcloning the corresponding polymerase chain reaction (PCR) fragments from the TSG101 gene together with PCR fragments from the pBI-EGFP plasmid (Clontech) into the pCXN2 mammalian expression vector ( ).

    Techniques: Generated, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Expressing, Transfection, Fluorescence, Microscopy

    Ectopically expressed intronless FHIT gene (cDNA) is spliced, generating a product equivalent to the endogenous aberrant mRNA in cancer cells. ( A ) Schematic representation of the FHIT pre-mRNA and the observed aberrant splicing in cancer cells (red line). The aligned sequences of the normal (full-length) and aberrant FHIT mRNAs reveal the skipping of exon 3 to exon 6. ( B ) Using normal cells (HMEC) and cancer cells (MCF-7), endogenous FHIT mRNAs (schematically shown) were analyzed by RT-PCR with the indicated primer sets (P25–P26, P25′–P26′). The full-length mRNA (black arrowhead) and aberrantly spliced mRNA (red arrowhead) are indicated. Black flags indicate the ORF (372–815) and the red line indicates the postulated mRNA re-splicing that skips the whole exon 3 to exon 6 region. ( C ) Cancer cells (MCF-7) were transfected with the FHIT-EGFP reporter plasmid (schematically shown) and the spliced products were analyzed by RT-PCR with the indicated primer sets (P25–P5, P25′–P6). The black and red arrowheads indicate the unspliced and spliced products, respectively. Sequence analysis mapped the spliced sites (red triangles) to the 5′ end of exon 3 and the 3′ end of exon 6 (data not shown).

    Journal: Nucleic Acids Research

    Article Title: Re-splicing of mature mRNA in cancer cells promotes activation of distant weak alternative splice sites

    doi: 10.1093/nar/gks520

    Figure Lengend Snippet: Ectopically expressed intronless FHIT gene (cDNA) is spliced, generating a product equivalent to the endogenous aberrant mRNA in cancer cells. ( A ) Schematic representation of the FHIT pre-mRNA and the observed aberrant splicing in cancer cells (red line). The aligned sequences of the normal (full-length) and aberrant FHIT mRNAs reveal the skipping of exon 3 to exon 6. ( B ) Using normal cells (HMEC) and cancer cells (MCF-7), endogenous FHIT mRNAs (schematically shown) were analyzed by RT-PCR with the indicated primer sets (P25–P26, P25′–P26′). The full-length mRNA (black arrowhead) and aberrantly spliced mRNA (red arrowhead) are indicated. Black flags indicate the ORF (372–815) and the red line indicates the postulated mRNA re-splicing that skips the whole exon 3 to exon 6 region. ( C ) Cancer cells (MCF-7) were transfected with the FHIT-EGFP reporter plasmid (schematically shown) and the spliced products were analyzed by RT-PCR with the indicated primer sets (P25–P5, P25′–P6). The black and red arrowheads indicate the unspliced and spliced products, respectively. Sequence analysis mapped the spliced sites (red triangles) to the 5′ end of exon 3 and the 3′ end of exon 6 (data not shown).

    Article Snippet: Construction of plasmids The expression plasmids for the TSG101-EGFP fusion proteins (TSG101-EGFP, TSG101-EGFP[+] and TSG101-EGFP[−]; C) were constructed by subcloning the corresponding polymerase chain reaction (PCR) fragments from the TSG101 gene together with PCR fragments from the pBI-EGFP plasmid (Clontech) into the pCXN2 mammalian expression vector ( ).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Transfection, Plasmid Preparation, Sequencing

    The lariat RNA containing TSG101 exon 2 to exon 9 was identified by sequencing. ( A ) Detection of the lariat exons by lariat-specific RT-PCR amplification across the branch point with the indicated primer sets (P23–P24, P23′–P24′). The RT-PCR signal remained in the RNase R-digested (+) RNA sample, revealing a closed lariat structure. ( B ) Sequencing analysis of the lariat-specific RT-PCR product. Electropherogram of the sequence containing the branch point (arrow), followed by the alternative 5′ splice site (arrow) in the lariat exons. ( C ) A sequence alignment of the PCR fragment (blue) and the TSG101 gene (black) reveals a 2′–5′ branched connection between the end of the alternative 5′ splice site ( GT ) and the branch point ( A ), which is located upstream from the alternative 3′ splice site ( AG ).

    Journal: Nucleic Acids Research

    Article Title: Re-splicing of mature mRNA in cancer cells promotes activation of distant weak alternative splice sites

    doi: 10.1093/nar/gks520

    Figure Lengend Snippet: The lariat RNA containing TSG101 exon 2 to exon 9 was identified by sequencing. ( A ) Detection of the lariat exons by lariat-specific RT-PCR amplification across the branch point with the indicated primer sets (P23–P24, P23′–P24′). The RT-PCR signal remained in the RNase R-digested (+) RNA sample, revealing a closed lariat structure. ( B ) Sequencing analysis of the lariat-specific RT-PCR product. Electropherogram of the sequence containing the branch point (arrow), followed by the alternative 5′ splice site (arrow) in the lariat exons. ( C ) A sequence alignment of the PCR fragment (blue) and the TSG101 gene (black) reveals a 2′–5′ branched connection between the end of the alternative 5′ splice site ( GT ) and the branch point ( A ), which is located upstream from the alternative 3′ splice site ( AG ).

    Article Snippet: Construction of plasmids The expression plasmids for the TSG101-EGFP fusion proteins (TSG101-EGFP, TSG101-EGFP[+] and TSG101-EGFP[−]; C) were constructed by subcloning the corresponding polymerase chain reaction (PCR) fragments from the TSG101 gene together with PCR fragments from the pBI-EGFP plasmid (Clontech) into the pCXN2 mammalian expression vector ( ).

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

    Detection of the lariat RNA consisting of only exons demonstrates the re-splicing of the endogenous TSG101 mRNA. ( A ) Schematic representation of the two postulated pathways leading to the generation of the aberrant mRNA, i.e. a conventional one-step direct aberrant splicing (right) and the proposed two-step process, including the re-splicing of the constitutively spliced mRNA (left). The pre-mRNA [1] and specific splicing products [2–5] from these two pathways were analyzed by RT-PCR with the indicated primers (arrows). ( B ) Detection of the specific splicing products [2–5] by RT-PCR using RNase R-digested (+) or RNase R-undigested (−) endogenous total RNA. The detected RT-PCR signals in the RNase R-digested sample (containing no linear RNAs) indicates an RNA species with either a 5′–2′ lariat or a 5′–3′ circular structure. However, the latter case was ruled out by the identification of a 5′–2′ branched structure ( Figure 4 A). Primers P19, P20, P21 and P22 anneal to introns 6, 7, 7 and 8, respectively. Primers P13/14 and P15/16 anneal to introns 7 and 8, respectively. Primers P11, P12, P17 and P18 anneal to exons 8, 10, 5 and 8, respectively. We used exactly the same PCR cycle numbers to amplify all these RNAs ( Supplementary Materials and Methods ).

    Journal: Nucleic Acids Research

    Article Title: Re-splicing of mature mRNA in cancer cells promotes activation of distant weak alternative splice sites

    doi: 10.1093/nar/gks520

    Figure Lengend Snippet: Detection of the lariat RNA consisting of only exons demonstrates the re-splicing of the endogenous TSG101 mRNA. ( A ) Schematic representation of the two postulated pathways leading to the generation of the aberrant mRNA, i.e. a conventional one-step direct aberrant splicing (right) and the proposed two-step process, including the re-splicing of the constitutively spliced mRNA (left). The pre-mRNA [1] and specific splicing products [2–5] from these two pathways were analyzed by RT-PCR with the indicated primers (arrows). ( B ) Detection of the specific splicing products [2–5] by RT-PCR using RNase R-digested (+) or RNase R-undigested (−) endogenous total RNA. The detected RT-PCR signals in the RNase R-digested sample (containing no linear RNAs) indicates an RNA species with either a 5′–2′ lariat or a 5′–3′ circular structure. However, the latter case was ruled out by the identification of a 5′–2′ branched structure ( Figure 4 A). Primers P19, P20, P21 and P22 anneal to introns 6, 7, 7 and 8, respectively. Primers P13/14 and P15/16 anneal to introns 7 and 8, respectively. Primers P11, P12, P17 and P18 anneal to exons 8, 10, 5 and 8, respectively. We used exactly the same PCR cycle numbers to amplify all these RNAs ( Supplementary Materials and Methods ).

    Article Snippet: Construction of plasmids The expression plasmids for the TSG101-EGFP fusion proteins (TSG101-EGFP, TSG101-EGFP[+] and TSG101-EGFP[−]; C) were constructed by subcloning the corresponding polymerase chain reaction (PCR) fragments from the TSG101 gene together with PCR fragments from the pBI-EGFP plasmid (Clontech) into the pCXN2 mammalian expression vector ( ).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction

    Real-time RT-PCR analyses of tHsc70 expression in the liver (a), skeletal muscle (b), lung (c), and heart (d) of P . sinensis after heat stress for 1, 2, and 4 h with 1-h adaptive recovery at 25 °C (Groups V, VI, and VII) or without adaptation

    Journal: Journal of Zhejiang University. Science. B

    Article Title: Identification of a heat shock cognate protein 70 gene in Chinese soft-shell turtle (Pelodiscus sinensis) and its expression profiles under thermal stress *

    doi: 10.1631/jzus.B1100309

    Figure Lengend Snippet: Real-time RT-PCR analyses of tHsc70 expression in the liver (a), skeletal muscle (b), lung (c), and heart (d) of P . sinensis after heat stress for 1, 2, and 4 h with 1-h adaptive recovery at 25 °C (Groups V, VI, and VII) or without adaptation

    Article Snippet: The polymerase chain reaction (PCR) fragment of tHsc70 was amplified using the primer pair Hsc70-dF and Hsc70-dR (Table ) on a thermal cycler S1000™ (Bio-Rad, Hercules, CA, USA).

    Techniques: Quantitative RT-PCR, Expressing

    Semi-quantitative RT-PCR analysis of tHsc70 expression in the liver, skeletal muscle, lung, and heart of P . sinensis

    Journal: Journal of Zhejiang University. Science. B

    Article Title: Identification of a heat shock cognate protein 70 gene in Chinese soft-shell turtle (Pelodiscus sinensis) and its expression profiles under thermal stress *

    doi: 10.1631/jzus.B1100309

    Figure Lengend Snippet: Semi-quantitative RT-PCR analysis of tHsc70 expression in the liver, skeletal muscle, lung, and heart of P . sinensis

    Article Snippet: The polymerase chain reaction (PCR) fragment of tHsc70 was amplified using the primer pair Hsc70-dF and Hsc70-dR (Table ) on a thermal cycler S1000™ (Bio-Rad, Hercules, CA, USA).

    Techniques: Quantitative RT-PCR, Expressing

    Relative expression profiles of down- (A) or up-regulated (B) monolignol genes in F . × ananassa cv. Elsanta . Total RNA was isolated from a single untreated fruit (wild type; WT), control fruits (pBI-intron), down-regulated fruits (pBI- FaCCRi , pBI- FaCADi , and pBI- FaPODi ) (A) , and up-regulated fruits (pBI- FaCCR , pBI- FaCAD , and pBI- FaPOD ) (B) . Expression levels of all samples were monitored by qRT-PCR with specific primers for target genes ( FaCCR . FaCAD , and FaPOD ) and an interspac er gene. The latter was used as an internal control for normalization. For each box-plot graph, one of the WT group was used as the reference (set to one) and each group contained five biological replicates. The Wilcoxon-Mann-Whitney U -test was used for a non-parametric comparison of two groups from pBI-intron and fruits infiltrated with different constructs. Values indicate statistically decreased and increased levels ( P

    Journal: Frontiers in Plant Science

    Article Title: FaPOD27 functions in the metabolism of polyphenols in strawberry fruit (Fragaria sp.)

    doi: 10.3389/fpls.2014.00518

    Figure Lengend Snippet: Relative expression profiles of down- (A) or up-regulated (B) monolignol genes in F . × ananassa cv. Elsanta . Total RNA was isolated from a single untreated fruit (wild type; WT), control fruits (pBI-intron), down-regulated fruits (pBI- FaCCRi , pBI- FaCADi , and pBI- FaPODi ) (A) , and up-regulated fruits (pBI- FaCCR , pBI- FaCAD , and pBI- FaPOD ) (B) . Expression levels of all samples were monitored by qRT-PCR with specific primers for target genes ( FaCCR . FaCAD , and FaPOD ) and an interspac er gene. The latter was used as an internal control for normalization. For each box-plot graph, one of the WT group was used as the reference (set to one) and each group contained five biological replicates. The Wilcoxon-Mann-Whitney U -test was used for a non-parametric comparison of two groups from pBI-intron and fruits infiltrated with different constructs. Values indicate statistically decreased and increased levels ( P

    Article Snippet: PCR fragments of the FaCCR and FaPOD cut with Bam HI and Sma I were subcloned into a Bam HI-Sma I cut pGEX-4T-1 vector (GE Healthcare, Munich, Germany), in frame with a coding region for an N-terminal GST (glutathione S-transferase) tag.

    Techniques: Expressing, Isolation, Quantitative RT-PCR, MANN-WHITNEY, Construct

    Relative expression profiles of monolignol biosynthesis genes in vegetative tissues, flowers, and fruit developmental stages of F . × ananassa cv. Elsanta . Total RNA was isolated from (A) fruit developmental stages at small green (SG), green white (GW), white (W), turning (T), and red (R) after pollination. (B) Expression levels of vegetative tissues (leaf; stem; runner; root), flower (F) and fruit developmental stages (SG, GW, W, T, and R) were monitored by qRT-PCR. FaCCR . FaCAD . FaPOD and FaPOD27 (Ring et al., 2013 ) were target genes. The white fruit was used as the reference with one for each graph. Values are mean ± SE of 5–6 replicates from two sets of cDNAs and are shown as relative changes.

    Journal: Frontiers in Plant Science

    Article Title: FaPOD27 functions in the metabolism of polyphenols in strawberry fruit (Fragaria sp.)

    doi: 10.3389/fpls.2014.00518

    Figure Lengend Snippet: Relative expression profiles of monolignol biosynthesis genes in vegetative tissues, flowers, and fruit developmental stages of F . × ananassa cv. Elsanta . Total RNA was isolated from (A) fruit developmental stages at small green (SG), green white (GW), white (W), turning (T), and red (R) after pollination. (B) Expression levels of vegetative tissues (leaf; stem; runner; root), flower (F) and fruit developmental stages (SG, GW, W, T, and R) were monitored by qRT-PCR. FaCCR . FaCAD . FaPOD and FaPOD27 (Ring et al., 2013 ) were target genes. The white fruit was used as the reference with one for each graph. Values are mean ± SE of 5–6 replicates from two sets of cDNAs and are shown as relative changes.

    Article Snippet: PCR fragments of the FaCCR and FaPOD cut with Bam HI and Sma I were subcloned into a Bam HI-Sma I cut pGEX-4T-1 vector (GE Healthcare, Munich, Germany), in frame with a coding region for an N-terminal GST (glutathione S-transferase) tag.

    Techniques: Expressing, Isolation, Quantitative RT-PCR

    Relative expression profiles of phenylpropanoid biosynthesis genes of F . × ananassa cv. Elsanta in response to Agrobacterium . Expression levels in control (gray column) and agroinfiltrated fruits (black column) were monitored by qRT-PCR at different time points (1, 3, 6, 12, 24, 48, and 96 h). FaPAL . FaCHS, FaCCR . FaCAD . FaPOD , and FaPOD27 were target genes. The control fruit (1 h) was used as the reference with one for each graph. Values are mean ± SE of 2–3 replicates from one fruit and are shown as relative changes.

    Journal: Frontiers in Plant Science

    Article Title: FaPOD27 functions in the metabolism of polyphenols in strawberry fruit (Fragaria sp.)

    doi: 10.3389/fpls.2014.00518

    Figure Lengend Snippet: Relative expression profiles of phenylpropanoid biosynthesis genes of F . × ananassa cv. Elsanta in response to Agrobacterium . Expression levels in control (gray column) and agroinfiltrated fruits (black column) were monitored by qRT-PCR at different time points (1, 3, 6, 12, 24, 48, and 96 h). FaPAL . FaCHS, FaCCR . FaCAD . FaPOD , and FaPOD27 were target genes. The control fruit (1 h) was used as the reference with one for each graph. Values are mean ± SE of 2–3 replicates from one fruit and are shown as relative changes.

    Article Snippet: PCR fragments of the FaCCR and FaPOD cut with Bam HI and Sma I were subcloned into a Bam HI-Sma I cut pGEX-4T-1 vector (GE Healthcare, Munich, Germany), in frame with a coding region for an N-terminal GST (glutathione S-transferase) tag.

    Techniques: Expressing, Quantitative RT-PCR

    MEF2C expression and PSL sensitivity of primary leukemic blast cells in four T-ALL patients (two ETP-ALL and two non-ETP ALL). (a) MEF2C expression levels determined by real-time quantitative-PCR. GAPDH was used as an internal control. (b) Viability of cells was counted after treatment with PSL (200 μM) and/or ABT-737 (10 nM). Annexin (-)/PI(-) cells were defined as viable cells.

    Journal: PLoS ONE

    Article Title: BCL2 Inhibitor (ABT-737): A Restorer of Prednisolone Sensitivity in Early T-Cell Precursor-Acute Lymphoblastic Leukemia with High MEF2C Expression?

    doi: 10.1371/journal.pone.0132926

    Figure Lengend Snippet: MEF2C expression and PSL sensitivity of primary leukemic blast cells in four T-ALL patients (two ETP-ALL and two non-ETP ALL). (a) MEF2C expression levels determined by real-time quantitative-PCR. GAPDH was used as an internal control. (b) Viability of cells was counted after treatment with PSL (200 μM) and/or ABT-737 (10 nM). Annexin (-)/PI(-) cells were defined as viable cells.

    Article Snippet: Establishment of BaF3 Cell Lines Expressing MEF2C Protein Retroviral constructs encoding MEF2C were generated by inserting the PCR fragment of MEF2C into the retroviral vector MSCVneo (Clontech, Mountain View, CA, USA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Expression levels of MEF2C and FLT3 in ETP-ALL vs. typical T-ALL blast cells. Comparison of the expression levels of MEF2C and FLT3 in ETP-ALL vs. typical T-ALL blast cells determined by real-time quantitative-PCR analysis.

    Journal: PLoS ONE

    Article Title: BCL2 Inhibitor (ABT-737): A Restorer of Prednisolone Sensitivity in Early T-Cell Precursor-Acute Lymphoblastic Leukemia with High MEF2C Expression?

    doi: 10.1371/journal.pone.0132926

    Figure Lengend Snippet: Expression levels of MEF2C and FLT3 in ETP-ALL vs. typical T-ALL blast cells. Comparison of the expression levels of MEF2C and FLT3 in ETP-ALL vs. typical T-ALL blast cells determined by real-time quantitative-PCR analysis.

    Article Snippet: Establishment of BaF3 Cell Lines Expressing MEF2C Protein Retroviral constructs encoding MEF2C were generated by inserting the PCR fragment of MEF2C into the retroviral vector MSCVneo (Clontech, Mountain View, CA, USA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Comparison of LOUCY and Jurkat leukemia cells. (a) Expression of MEF2C and β-actin by western blot analyses. (b), (c) Cell growth inhibition of leukemia cells assessed by WST assay with serial concentrations of prednisolone (PSL) or ABT-737. (d) q-PCR analysis of BCL2 of two cell lines 12 hours after treatment of ABT-737 with the concentration of 10 or 100 nM. The bar indicates the mean±SE of two independent experiments in triplicate (e) Cell growth inhibition with serial concentrations of prednisolone (PSL) combined with ABT-737 (10 nM). (f) Calculation of the IC50 of PSL, following treatment of leukemic cells with either PSL, or a combination of PSL and ABT-737. Calculated combination Index of less than 1.0 was considered as a synergistic effect. (g), (h) Apoptosis study. Annexin V-positive cells were counted as apoptotic. Flow cytometry data in (g) and the mean±SE of three independent experiments in (h).

    Journal: PLoS ONE

    Article Title: BCL2 Inhibitor (ABT-737): A Restorer of Prednisolone Sensitivity in Early T-Cell Precursor-Acute Lymphoblastic Leukemia with High MEF2C Expression?

    doi: 10.1371/journal.pone.0132926

    Figure Lengend Snippet: Comparison of LOUCY and Jurkat leukemia cells. (a) Expression of MEF2C and β-actin by western blot analyses. (b), (c) Cell growth inhibition of leukemia cells assessed by WST assay with serial concentrations of prednisolone (PSL) or ABT-737. (d) q-PCR analysis of BCL2 of two cell lines 12 hours after treatment of ABT-737 with the concentration of 10 or 100 nM. The bar indicates the mean±SE of two independent experiments in triplicate (e) Cell growth inhibition with serial concentrations of prednisolone (PSL) combined with ABT-737 (10 nM). (f) Calculation of the IC50 of PSL, following treatment of leukemic cells with either PSL, or a combination of PSL and ABT-737. Calculated combination Index of less than 1.0 was considered as a synergistic effect. (g), (h) Apoptosis study. Annexin V-positive cells were counted as apoptotic. Flow cytometry data in (g) and the mean±SE of three independent experiments in (h).

    Article Snippet: Establishment of BaF3 Cell Lines Expressing MEF2C Protein Retroviral constructs encoding MEF2C were generated by inserting the PCR fragment of MEF2C into the retroviral vector MSCVneo (Clontech, Mountain View, CA, USA).

    Techniques: Expressing, Western Blot, Inhibition, WST Assay, Polymerase Chain Reaction, Concentration Assay, Flow Cytometry, Cytometry

    Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative PCR (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from MiSeq analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P

    Journal: Genes & Development

    Article Title: Mutant TRP53 exerts a target gene-selective dominant-negative effect to drive tumor development

    doi: 10.1101/gad.314286.118

    Figure Lengend Snippet: Overexpression of mutant TRP53 proteins accelerates lymphoma development in an Eμ-Myc;Trp53 +/+ background and relieves selective pressure for mutation of endogenous Trp53 genes. ( A ) Kaplan-Meier survival curves for mice reconstituted with Eμ-Myc;Trp53 +/+ HSPCs comparing empty vector control (pMIG), CRISPR/Cas9 Trp53 knockout , and each mutant TRP53 protein (V170M, I192S, G280, R246Q, and R270H). P -values were determined by log rank (Mantel-Cox) test. ( B ) Selected TRP53 protein immunohistochemistry in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments (mice #88 and #541 plus control mouse #53). ( C ) Endogenous Trp53 allele copy number in lymphomas from the Eμ-Myc hematopoietic reconstitution experiments as determined by genomic DNA quantitative PCR (pMIG/control: #53; V170M: #55, #66, and #541; G280: #68 and #81; I192S: #72 and #546; R246Q: #97 and #98; R270H: #80, #543, and #544). Primary cells from Trp53 −/− and Trp53 +/− mice were used as controls. Data from MiSeq analysis throughout the coding region of the DNA-binding domain (exons 4–10) are indicated. (wt) Wild-type sequence. Data represent mean ± SEM. (*) P

    Article Snippet: PCR fragments were sequenced using the MiSeq platform (Illumina).

    Techniques: Over Expression, Mutagenesis, Mouse Assay, Plasmid Preparation, CRISPR, Knock-Out, Immunohistochemistry, Real-time Polymerase Chain Reaction, Binding Assay, Sequencing