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  • 95
    Millipore complementary dna cdna synthesis kits
    Complementary Dna Cdna Synthesis Kits, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher complementary dna cdna synthesis kit
    Gene expression profiles in GANT61-treated SKOV3 cells. ( A ) Identification of differentially expressed genes (DEGs) in GANT61-treated SKOV3 cells. Cells were treated with GANT61 (20 µM) or DMSO for 60 hr, and total <t>RNA</t> was extracted as described in Methods . Changes in gene expression were determined by <t>cDNA</t> microarray gene profiling using the Illumina Human HT expression BeadChip V4 (Illumina Inc., San Diego, CA). Differential expressions for the DEGs are shown. ( B ) The top 11 canonical signaling pathways influenced by inhibition of Gli1/Gli2 function in SKOV3 cells. The top 11 canonical signaling pathways, determined by IPA, that were significantly up-regulated or down-regulated by GANT61 treatment in SKOV3 cells, are shown. The 412 DEGs were mapped to the IPA-defined network. The significant P -values that determine the probability that the association between the genes in the dataset and the canonical pathway is by chance alone were calculated by Fisher's exact test, and are expressed as −log ( P -value). ( C ) Heat map of DEGs in the focal adhesion signal pathway in GANT61-treated SKOV3 cells. The heat map shows that 17 genes were significantly differentially expressed, including seven up-regulated genes and 12 down-regulated genes, in the GANT61-treated compared to control SKOV3 cells. ( D ) Selected DEGs from cDNA array gene expression profiling analyzed by real-time PCR. SKOV3 cells were treated with DMSO or GANT61 for the indicated times, total RNA was extracted for real-time PCR as described in Methods using the primer sets in Table 1 . The data represent the mean ± SD of three determinations, and GAPDH was used to normalize the relative mRNA levels.
    Complementary Dna Cdna Synthesis Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega complementary dna cdna synthesis kit
    Tissue expression of hMSH6 <t>mRNA.</t> A human multiple-tissue Northern blot (CLONTECH) was probed with the 32 P-labeled complete <t>cDNA</t> clone of hMSH6 according to the manufacturer’s protocol. Colon refers to mucosal lining; p.b. leukocyte refers to peripheral blood leukocyte. The hMSH6 transcript is indicated by the arrow and corresponds to ≈4.5 kb. The amount of RNA loaded in each lane was adjusted to comparable levels as judged spectrophotometrically and by the levels of actin present (CLONTECH).
    Complementary Dna Cdna Synthesis Kit, supplied by Promega, used in various techniques. Bioz Stars score: 88/100, based on 193 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bioneer Corporation complementary dna cdna synthesis kit
    Tissue expression of hMSH6 <t>mRNA.</t> A human multiple-tissue Northern blot (CLONTECH) was probed with the 32 P-labeled complete <t>cDNA</t> clone of hMSH6 according to the manufacturer’s protocol. Colon refers to mucosal lining; p.b. leukocyte refers to peripheral blood leukocyte. The hMSH6 transcript is indicated by the arrow and corresponds to ≈4.5 kb. The amount of RNA loaded in each lane was adjusted to comparable levels as judged spectrophotometrically and by the levels of actin present (CLONTECH).
    Complementary Dna Cdna Synthesis Kit, supplied by Bioneer Corporation, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad complementary dna cdna synthesis kit
    Immunohistologic analysis of TELi. (A–B): H E staining of native liver and TELi. An asterisk is placed near a blood vessel containing erythrocytes in TELi. White arrows indicate incompletely degraded scaffold material. (C): Immunofluorescence staining for albumin (red) and CK8 (cholangiocyte and periductular hepatocyte marker, green) in native liver and TELi. Arrowhead shows double positive cells. (D): Immunofluorescence staining for HNF4α (hepatocyte specific transcription factor, red) and CK8 (green). Nuclei stained with 4′,6‐diamidino‐2‐phenylindole (blue). (E): Reverse transcription‐polymerase chain reaction for albumin expression. <t>cDNA</t> prepared from whole liver <t>RNA.</t> Scale bars = 50 μm (A) , 25 μm (B–D) . Data represent more than three independent experiments. Abbreviations: H E, hematoxylin and eosin; L, ladder; NC, negative control (water); PC, positive control; TELi, tissue‐engineered liver.
    Complementary Dna Cdna Synthesis Kit, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa complementary dna cdna synthesis kit
    Effects of DCA on mRNA and protein levels of OCTN2 in cancer cell lines. (A), analysis of mRNA levels of OCTN2 in DCA-treated cancer cell lines. After treatment with 0.5 and 1 μM DCA, total <t>RNA</t> in cancer cells was purified by TRIzol reagent for <t>cDNA</t> synthesis and the mRNA levels of OCTN2 were analyzed by Quantitative RT-PCR. (B), analysis of protein levels of OCTN2 in DCA-treated cancer cell lines. Total proteins were extracted and protein levels of OCTN2 were analyzed by western blotting. The relative expression of mRNA and protein was normalized to β-actin. Data presented represent the mean ± S.D. of three independent experiments. Significant difference from control group of 0.1% DMSO is denoted with asterisks (*, p
    Complementary Dna Cdna Synthesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche cdna synthesis kit
    Messenger <t>RNA</t> expression of TNC receptors of MCF-7 cells after TGF-β and TNC treatments. ( a ) mRNA was extracted from cells co-stimulated with TNC (10 μg/ml) and TGF-β1 (5 ng/ml) for synthesis of <t>cDNA</t> and performance of real-time PCR determining the cycle threshold ( C t ) values for each subunit. Differences in C t values of TNC receptor mRNAs from that of GAPDH mRNA in TGF-β1/TNC-treated cells are shown using qPCR. C t values of α7/8/9 and β3 integrin subunits were very high, indicating only limited expression. Calibration curves were obtained using each target gene in the samples. ( b ) Changes of receptor mRNA expression for αv/2 and β1/6 after treatment with TGF-β1 and/or TNC. The results were normalized to GAPDH expression. Integrin αv showed significant increase after TGF-β1/TNC treatment and β6 was dramatically increased after TGF-β1 addition, with or without TNC (mean±s.d.; * P
    Cdna Synthesis Kit, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 1391 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Quanta Biosciences cdna synthesis kit
    Messenger <t>RNA</t> expression of TNC receptors of MCF-7 cells after TGF-β and TNC treatments. ( a ) mRNA was extracted from cells co-stimulated with TNC (10 μg/ml) and TGF-β1 (5 ng/ml) for synthesis of <t>cDNA</t> and performance of real-time PCR determining the cycle threshold ( C t ) values for each subunit. Differences in C t values of TNC receptor mRNAs from that of GAPDH mRNA in TGF-β1/TNC-treated cells are shown using qPCR. C t values of α7/8/9 and β3 integrin subunits were very high, indicating only limited expression. Calibration curves were obtained using each target gene in the samples. ( b ) Changes of receptor mRNA expression for αv/2 and β1/6 after treatment with TGF-β1 and/or TNC. The results were normalized to GAPDH expression. Integrin αv showed significant increase after TGF-β1/TNC treatment and β6 was dramatically increased after TGF-β1 addition, with or without TNC (mean±s.d.; * P
    Cdna Synthesis Kit, supplied by Quanta Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 332 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Quanta Biosciences superscript complementary dna cdna synthesis kit
    Messenger <t>RNA</t> expression of TNC receptors of MCF-7 cells after TGF-β and TNC treatments. ( a ) mRNA was extracted from cells co-stimulated with TNC (10 μg/ml) and TGF-β1 (5 ng/ml) for synthesis of <t>cDNA</t> and performance of real-time PCR determining the cycle threshold ( C t ) values for each subunit. Differences in C t values of TNC receptor mRNAs from that of GAPDH mRNA in TGF-β1/TNC-treated cells are shown using qPCR. C t values of α7/8/9 and β3 integrin subunits were very high, indicating only limited expression. Calibration curves were obtained using each target gene in the samples. ( b ) Changes of receptor mRNA expression for αv/2 and β1/6 after treatment with TGF-β1 and/or TNC. The results were normalized to GAPDH expression. Integrin αv showed significant increase after TGF-β1/TNC treatment and β6 was dramatically increased after TGF-β1 addition, with or without TNC (mean±s.d.; * P
    Superscript Complementary Dna Cdna Synthesis Kit, supplied by Quanta Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher complementary dna cdna synthesis supermix kit
    Messenger <t>RNA</t> expression of TNC receptors of MCF-7 cells after TGF-β and TNC treatments. ( a ) mRNA was extracted from cells co-stimulated with TNC (10 μg/ml) and TGF-β1 (5 ng/ml) for synthesis of <t>cDNA</t> and performance of real-time PCR determining the cycle threshold ( C t ) values for each subunit. Differences in C t values of TNC receptor mRNAs from that of GAPDH mRNA in TGF-β1/TNC-treated cells are shown using qPCR. C t values of α7/8/9 and β3 integrin subunits were very high, indicating only limited expression. Calibration curves were obtained using each target gene in the samples. ( b ) Changes of receptor mRNA expression for αv/2 and β1/6 after treatment with TGF-β1 and/or TNC. The results were normalized to GAPDH expression. Integrin αv showed significant increase after TGF-β1/TNC treatment and β6 was dramatically increased after TGF-β1 addition, with or without TNC (mean±s.d.; * P
    Complementary Dna Cdna Synthesis Supermix Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TransGen biotech co cdna synthesis kit
    Messenger <t>RNA</t> expression of TNC receptors of MCF-7 cells after TGF-β and TNC treatments. ( a ) mRNA was extracted from cells co-stimulated with TNC (10 μg/ml) and TGF-β1 (5 ng/ml) for synthesis of <t>cDNA</t> and performance of real-time PCR determining the cycle threshold ( C t ) values for each subunit. Differences in C t values of TNC receptor mRNAs from that of GAPDH mRNA in TGF-β1/TNC-treated cells are shown using qPCR. C t values of α7/8/9 and β3 integrin subunits were very high, indicating only limited expression. Calibration curves were obtained using each target gene in the samples. ( b ) Changes of receptor mRNA expression for αv/2 and β1/6 after treatment with TGF-β1 and/or TNC. The results were normalized to GAPDH expression. Integrin αv showed significant increase after TGF-β1/TNC treatment and β6 was dramatically increased after TGF-β1 addition, with or without TNC (mean±s.d.; * P
    Cdna Synthesis Kit, supplied by TransGen biotech co, used in various techniques. Bioz Stars score: 93/100, based on 179 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene cdna synthesis kit
    The nucleotide and deduced amino acid sequences of Fus p 4.0101 (GenBank accession no. KF151224). Numbers to the right are the positions of the nucleotides and the deduced residues of the sequences. The stop codon TAA is denoted with an asterisk (*). The potential N-glycosylation site is indicated in bold letters and boxed. Eight conserved amino acid residues that are involved in the catalysis and the substrate binding of transaldolases are shaded and in bold letters. Nucleotide sequences in grey correspond to primer sequences synthesized for <t>PCR</t> experiments in the <t>cDNA</t> cloning of Fus p 4.0101 as described in the Materials and Methods. The sequences corresponding to primers Fu-TAase-f and Fu-TAase-r used in the preparation of rFus p 4.0101 are boxed.
    Cdna Synthesis Kit, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 1002 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies cdna synthesis kit
    Identification of Oplr16 as a pluripotent lncRNA. ( A ) Identification of Oplr16 as a pluripotency-associated lncRNA by integrating RNA-seq and CRIST-seq. RNA-seq was used to identify RNAs that are differentially expressed between fibroblasts and iPSCs collected in the process of reprogramming. In CRIST-seq, the Oct4 promoter was targeted by the expression of the Cas9 Oct4 -gRNA. LncRNAs interacting with Oct4 promoter were reverse transcribed in situ into <t>cDNA</t> with biotin-dCTP. After immunoprecipitation, biotin-cDNAs were purified by streptavidin beads for <t>Illumina</t> sequencing. Both sets of databases were integrated to identify the pluripotency-associated lncRNA Oplr16 . ( B ) Reactivation of Oplr16 in reprogramming. Fibroblasts were transfected with a Oct4-Sox2-Kilf4-c-Myc (OSKM) lentivirus. Cells were collected at various stages of reprogramming and expression of Oplr16 was measured by qPCR. E14: mouse embryonic pluripotent stem cell line used as a positive control; iPSC: induced pluripotent stem cells; non-iPSC: un-reprogrammed cells that express four OSKM factors, but fail to complete reprogramming; FBC: fibroblasts. β-Actin was used as the PCR control. ** P
    Cdna Synthesis Kit, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 93/100, based on 732 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen complementary dna cdna synthesis kit
    Identification of Oplr16 as a pluripotent lncRNA. ( A ) Identification of Oplr16 as a pluripotency-associated lncRNA by integrating RNA-seq and CRIST-seq. RNA-seq was used to identify RNAs that are differentially expressed between fibroblasts and iPSCs collected in the process of reprogramming. In CRIST-seq, the Oct4 promoter was targeted by the expression of the Cas9 Oct4 -gRNA. LncRNAs interacting with Oct4 promoter were reverse transcribed in situ into <t>cDNA</t> with biotin-dCTP. After immunoprecipitation, biotin-cDNAs were purified by streptavidin beads for <t>Illumina</t> sequencing. Both sets of databases were integrated to identify the pluripotency-associated lncRNA Oplr16 . ( B ) Reactivation of Oplr16 in reprogramming. Fibroblasts were transfected with a Oct4-Sox2-Kilf4-c-Myc (OSKM) lentivirus. Cells were collected at various stages of reprogramming and expression of Oplr16 was measured by qPCR. E14: mouse embryonic pluripotent stem cell line used as a positive control; iPSC: induced pluripotent stem cells; non-iPSC: un-reprogrammed cells that express four OSKM factors, but fail to complete reprogramming; FBC: fibroblasts. β-Actin was used as the PCR control. ** P
    Complementary Dna Cdna Synthesis Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega goscript complementary dna cdna synthesis kit
    Identification of Oplr16 as a pluripotent lncRNA. ( A ) Identification of Oplr16 as a pluripotency-associated lncRNA by integrating RNA-seq and CRIST-seq. RNA-seq was used to identify RNAs that are differentially expressed between fibroblasts and iPSCs collected in the process of reprogramming. In CRIST-seq, the Oct4 promoter was targeted by the expression of the Cas9 Oct4 -gRNA. LncRNAs interacting with Oct4 promoter were reverse transcribed in situ into <t>cDNA</t> with biotin-dCTP. After immunoprecipitation, biotin-cDNAs were purified by streptavidin beads for <t>Illumina</t> sequencing. Both sets of databases were integrated to identify the pluripotency-associated lncRNA Oplr16 . ( B ) Reactivation of Oplr16 in reprogramming. Fibroblasts were transfected with a Oct4-Sox2-Kilf4-c-Myc (OSKM) lentivirus. Cells were collected at various stages of reprogramming and expression of Oplr16 was measured by qPCR. E14: mouse embryonic pluripotent stem cell line used as a positive control; iPSC: induced pluripotent stem cells; non-iPSC: un-reprogrammed cells that express four OSKM factors, but fail to complete reprogramming; FBC: fibroblasts. β-Actin was used as the PCR control. ** P
    Goscript Complementary Dna Cdna Synthesis Kit, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Meridian Life Science cdna synthesis kit
    Identification of Oplr16 as a pluripotent lncRNA. ( A ) Identification of Oplr16 as a pluripotency-associated lncRNA by integrating RNA-seq and CRIST-seq. RNA-seq was used to identify RNAs that are differentially expressed between fibroblasts and iPSCs collected in the process of reprogramming. In CRIST-seq, the Oct4 promoter was targeted by the expression of the Cas9 Oct4 -gRNA. LncRNAs interacting with Oct4 promoter were reverse transcribed in situ into <t>cDNA</t> with biotin-dCTP. After immunoprecipitation, biotin-cDNAs were purified by streptavidin beads for <t>Illumina</t> sequencing. Both sets of databases were integrated to identify the pluripotency-associated lncRNA Oplr16 . ( B ) Reactivation of Oplr16 in reprogramming. Fibroblasts were transfected with a Oct4-Sox2-Kilf4-c-Myc (OSKM) lentivirus. Cells were collected at various stages of reprogramming and expression of Oplr16 was measured by qPCR. E14: mouse embryonic pluripotent stem cell line used as a positive control; iPSC: induced pluripotent stem cells; non-iPSC: un-reprogrammed cells that express four OSKM factors, but fail to complete reprogramming; FBC: fibroblasts. β-Actin was used as the PCR control. ** P
    Cdna Synthesis Kit, supplied by Meridian Life Science, used in various techniques. Bioz Stars score: 92/100, based on 482 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Quanta Biosciences qscript cdna synthesis kit
    Identification of Oplr16 as a pluripotent lncRNA. ( A ) Identification of Oplr16 as a pluripotency-associated lncRNA by integrating RNA-seq and CRIST-seq. RNA-seq was used to identify RNAs that are differentially expressed between fibroblasts and iPSCs collected in the process of reprogramming. In CRIST-seq, the Oct4 promoter was targeted by the expression of the Cas9 Oct4 -gRNA. LncRNAs interacting with Oct4 promoter were reverse transcribed in situ into <t>cDNA</t> with biotin-dCTP. After immunoprecipitation, biotin-cDNAs were purified by streptavidin beads for <t>Illumina</t> sequencing. Both sets of databases were integrated to identify the pluripotency-associated lncRNA Oplr16 . ( B ) Reactivation of Oplr16 in reprogramming. Fibroblasts were transfected with a Oct4-Sox2-Kilf4-c-Myc (OSKM) lentivirus. Cells were collected at various stages of reprogramming and expression of Oplr16 was measured by qPCR. E14: mouse embryonic pluripotent stem cell line used as a positive control; iPSC: induced pluripotent stem cells; non-iPSC: un-reprogrammed cells that express four OSKM factors, but fail to complete reprogramming; FBC: fibroblasts. β-Actin was used as the PCR control. ** P
    Qscript Cdna Synthesis Kit, supplied by Quanta Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 4783 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Gene expression profiles in GANT61-treated SKOV3 cells. ( A ) Identification of differentially expressed genes (DEGs) in GANT61-treated SKOV3 cells. Cells were treated with GANT61 (20 µM) or DMSO for 60 hr, and total RNA was extracted as described in Methods . Changes in gene expression were determined by cDNA microarray gene profiling using the Illumina Human HT expression BeadChip V4 (Illumina Inc., San Diego, CA). Differential expressions for the DEGs are shown. ( B ) The top 11 canonical signaling pathways influenced by inhibition of Gli1/Gli2 function in SKOV3 cells. The top 11 canonical signaling pathways, determined by IPA, that were significantly up-regulated or down-regulated by GANT61 treatment in SKOV3 cells, are shown. The 412 DEGs were mapped to the IPA-defined network. The significant P -values that determine the probability that the association between the genes in the dataset and the canonical pathway is by chance alone were calculated by Fisher's exact test, and are expressed as −log ( P -value). ( C ) Heat map of DEGs in the focal adhesion signal pathway in GANT61-treated SKOV3 cells. The heat map shows that 17 genes were significantly differentially expressed, including seven up-regulated genes and 12 down-regulated genes, in the GANT61-treated compared to control SKOV3 cells. ( D ) Selected DEGs from cDNA array gene expression profiling analyzed by real-time PCR. SKOV3 cells were treated with DMSO or GANT61 for the indicated times, total RNA was extracted for real-time PCR as described in Methods using the primer sets in Table 1 . The data represent the mean ± SD of three determinations, and GAPDH was used to normalize the relative mRNA levels.

    Journal: PLoS ONE

    Article Title: Down-Regulation of Gli Transcription Factor Leads to the Inhibition of Migration and Invasion of Ovarian Cancer Cells via Integrin ?4-Mediated FAK Signaling

    doi: 10.1371/journal.pone.0088386

    Figure Lengend Snippet: Gene expression profiles in GANT61-treated SKOV3 cells. ( A ) Identification of differentially expressed genes (DEGs) in GANT61-treated SKOV3 cells. Cells were treated with GANT61 (20 µM) or DMSO for 60 hr, and total RNA was extracted as described in Methods . Changes in gene expression were determined by cDNA microarray gene profiling using the Illumina Human HT expression BeadChip V4 (Illumina Inc., San Diego, CA). Differential expressions for the DEGs are shown. ( B ) The top 11 canonical signaling pathways influenced by inhibition of Gli1/Gli2 function in SKOV3 cells. The top 11 canonical signaling pathways, determined by IPA, that were significantly up-regulated or down-regulated by GANT61 treatment in SKOV3 cells, are shown. The 412 DEGs were mapped to the IPA-defined network. The significant P -values that determine the probability that the association between the genes in the dataset and the canonical pathway is by chance alone were calculated by Fisher's exact test, and are expressed as −log ( P -value). ( C ) Heat map of DEGs in the focal adhesion signal pathway in GANT61-treated SKOV3 cells. The heat map shows that 17 genes were significantly differentially expressed, including seven up-regulated genes and 12 down-regulated genes, in the GANT61-treated compared to control SKOV3 cells. ( D ) Selected DEGs from cDNA array gene expression profiling analyzed by real-time PCR. SKOV3 cells were treated with DMSO or GANT61 for the indicated times, total RNA was extracted for real-time PCR as described in Methods using the primer sets in Table 1 . The data represent the mean ± SD of three determinations, and GAPDH was used to normalize the relative mRNA levels.

    Article Snippet: Real-time PCR Total RNA (1 µg) was employed to prepare cDNA via reverse transcription using cDNA synthesis kit (Invitrogen) according to manufacturer's instructions and analyzed using an Applied Biosystems 7500 PCR Detection System (Applied Biosystems Inc.).

    Techniques: Expressing, Microarray, Inhibition, Indirect Immunoperoxidase Assay, Real-time Polymerase Chain Reaction

    Tissue expression of hMSH6 mRNA. A human multiple-tissue Northern blot (CLONTECH) was probed with the 32 P-labeled complete cDNA clone of hMSH6 according to the manufacturer’s protocol. Colon refers to mucosal lining; p.b. leukocyte refers to peripheral blood leukocyte. The hMSH6 transcript is indicated by the arrow and corresponds to ≈4.5 kb. The amount of RNA loaded in each lane was adjusted to comparable levels as judged spectrophotometrically and by the levels of actin present (CLONTECH).

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

    Article Title: hMSH2 forms specific mispair-binding complexes with hMSH3 and hMSH6

    doi:

    Figure Lengend Snippet: Tissue expression of hMSH6 mRNA. A human multiple-tissue Northern blot (CLONTECH) was probed with the 32 P-labeled complete cDNA clone of hMSH6 according to the manufacturer’s protocol. Colon refers to mucosal lining; p.b. leukocyte refers to peripheral blood leukocyte. The hMSH6 transcript is indicated by the arrow and corresponds to ≈4.5 kb. The amount of RNA loaded in each lane was adjusted to comparable levels as judged spectrophotometrically and by the levels of actin present (CLONTECH).

    Article Snippet: Single-strand, random-primed cDNA was prepared from human testis poly(A) mRNA (CLONTECH) using a Promega cDNA synthesis kit.

    Techniques: Expressing, Northern Blot, Labeling

    Organization of hMSH6 genomic locus and sequence of the intron region flanking each MSH6 exon. Boxes containing numbers 1 through 10 indicate the individual MSH6 exons and their relative sizes. The size of each exon is given below each exon and the size of each intron is given above the region between each pair of exons. The sizes of exons 1 and 10 are the sizes of the mRNA sequence upstream and downstream of the first and last introns, respectively; these sizes were calculated from the longest MSH6 cDNA sequence available. The first 20 nucleotides of each intron sequence up to the intron-exon junction is given in uppercase letters except for the 3′ side of the last intron, where additional sequence is given. The first 10 nucleotides of each exon sequence up to the intron-exon junction is given in lowercase letters. In addition, the sequence of the 5′ and 3′ ends of the cDNA, minus the poly(A) sequence, is also given in lowercase letters. The numbers in parentheses between intron sequences are the nucleotide coordinates of the exon sequences or cDNA sequences, assuming the A of the ATG is nucleotide 1. (Additional intron sequence including the complete sequence of some introns has been determined in all cases and is available on request from M.F.K. and R.K.)

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

    Article Title: hMSH2 forms specific mispair-binding complexes with hMSH3 and hMSH6

    doi:

    Figure Lengend Snippet: Organization of hMSH6 genomic locus and sequence of the intron region flanking each MSH6 exon. Boxes containing numbers 1 through 10 indicate the individual MSH6 exons and their relative sizes. The size of each exon is given below each exon and the size of each intron is given above the region between each pair of exons. The sizes of exons 1 and 10 are the sizes of the mRNA sequence upstream and downstream of the first and last introns, respectively; these sizes were calculated from the longest MSH6 cDNA sequence available. The first 20 nucleotides of each intron sequence up to the intron-exon junction is given in uppercase letters except for the 3′ side of the last intron, where additional sequence is given. The first 10 nucleotides of each exon sequence up to the intron-exon junction is given in lowercase letters. In addition, the sequence of the 5′ and 3′ ends of the cDNA, minus the poly(A) sequence, is also given in lowercase letters. The numbers in parentheses between intron sequences are the nucleotide coordinates of the exon sequences or cDNA sequences, assuming the A of the ATG is nucleotide 1. (Additional intron sequence including the complete sequence of some introns has been determined in all cases and is available on request from M.F.K. and R.K.)

    Article Snippet: Single-strand, random-primed cDNA was prepared from human testis poly(A) mRNA (CLONTECH) using a Promega cDNA synthesis kit.

    Techniques: Sequencing

    Immunohistologic analysis of TELi. (A–B): H E staining of native liver and TELi. An asterisk is placed near a blood vessel containing erythrocytes in TELi. White arrows indicate incompletely degraded scaffold material. (C): Immunofluorescence staining for albumin (red) and CK8 (cholangiocyte and periductular hepatocyte marker, green) in native liver and TELi. Arrowhead shows double positive cells. (D): Immunofluorescence staining for HNF4α (hepatocyte specific transcription factor, red) and CK8 (green). Nuclei stained with 4′,6‐diamidino‐2‐phenylindole (blue). (E): Reverse transcription‐polymerase chain reaction for albumin expression. cDNA prepared from whole liver RNA. Scale bars = 50 μm (A) , 25 μm (B–D) . Data represent more than three independent experiments. Abbreviations: H E, hematoxylin and eosin; L, ladder; NC, negative control (water); PC, positive control; TELi, tissue‐engineered liver.

    Journal: Stem Cells Translational Medicine

    Article Title: Functional Human and Murine Tissue‐Engineered Liver Is Generated from Adult Stem/Progenitor Cells

    doi: 10.5966/sctm.2016-0205

    Figure Lengend Snippet: Immunohistologic analysis of TELi. (A–B): H E staining of native liver and TELi. An asterisk is placed near a blood vessel containing erythrocytes in TELi. White arrows indicate incompletely degraded scaffold material. (C): Immunofluorescence staining for albumin (red) and CK8 (cholangiocyte and periductular hepatocyte marker, green) in native liver and TELi. Arrowhead shows double positive cells. (D): Immunofluorescence staining for HNF4α (hepatocyte specific transcription factor, red) and CK8 (green). Nuclei stained with 4′,6‐diamidino‐2‐phenylindole (blue). (E): Reverse transcription‐polymerase chain reaction for albumin expression. cDNA prepared from whole liver RNA. Scale bars = 50 μm (A) , 25 μm (B–D) . Data represent more than three independent experiments. Abbreviations: H E, hematoxylin and eosin; L, ladder; NC, negative control (water); PC, positive control; TELi, tissue‐engineered liver.

    Article Snippet: Reverse Transcription‐Polymerase Chain Reaction RNA was isolated from frozen TELi with the Qiagen RNA purification kit. cDNA was prepared from 1000 ng of RNA using Bio‐Rad cDNA synthesis kit (Bio‐Rad, Hercules, CA, http://www.bio-rad.com ).

    Techniques: Staining, Immunofluorescence, Marker, Reverse Transcription Polymerase Chain Reaction, Expressing, Negative Control, Positive Control

    Effects of DCA on mRNA and protein levels of OCTN2 in cancer cell lines. (A), analysis of mRNA levels of OCTN2 in DCA-treated cancer cell lines. After treatment with 0.5 and 1 μM DCA, total RNA in cancer cells was purified by TRIzol reagent for cDNA synthesis and the mRNA levels of OCTN2 were analyzed by Quantitative RT-PCR. (B), analysis of protein levels of OCTN2 in DCA-treated cancer cell lines. Total proteins were extracted and protein levels of OCTN2 were analyzed by western blotting. The relative expression of mRNA and protein was normalized to β-actin. Data presented represent the mean ± S.D. of three independent experiments. Significant difference from control group of 0.1% DMSO is denoted with asterisks (*, p

    Journal: PLoS ONE

    Article Title: Different Involvement of Promoter Methylation in the Expression of Organic Cation/Carnitine Transporter 2 (OCTN2) in Cancer Cell Lines

    doi: 10.1371/journal.pone.0076474

    Figure Lengend Snippet: Effects of DCA on mRNA and protein levels of OCTN2 in cancer cell lines. (A), analysis of mRNA levels of OCTN2 in DCA-treated cancer cell lines. After treatment with 0.5 and 1 μM DCA, total RNA in cancer cells was purified by TRIzol reagent for cDNA synthesis and the mRNA levels of OCTN2 were analyzed by Quantitative RT-PCR. (B), analysis of protein levels of OCTN2 in DCA-treated cancer cell lines. Total proteins were extracted and protein levels of OCTN2 were analyzed by western blotting. The relative expression of mRNA and protein was normalized to β-actin. Data presented represent the mean ± S.D. of three independent experiments. Significant difference from control group of 0.1% DMSO is denoted with asterisks (*, p

    Article Snippet: RNA purification, cDNA Synthesis and Quantitative Real-time PCR Total cellular RNA was isolated from the treated cells using TRIzol reagent. cDNA was synthesized from 1 μg of total RNA using the first cDNA synthesis kit (Takara, Dalian).

    Techniques: Purification, Quantitative RT-PCR, Western Blot, Expressing

    Expression of OCTN2 in cancer cell lines. (A), analysis of mRNA levels of OCTN2 in cancer cell lines. Total RNA in cancer cells was purified by TRIzol reagent for cDNA synthesis and the mRNA levels of OCTN2 were analyzed by Quantitative RT-PCR. (B), analysis of protein levels of OCTN2 in cancer cells. Total proteins were extracted and protein levels of OCTN2 were analyzed by western blotting. The relative expression of mRNA and protein was normalized to β-actin. The expression of HepG2 was set as 1. All of mRNA and protein analysis were performed three times. Significant difference from HepG2 or LS174T cell is denoted with asterisk (*, p

    Journal: PLoS ONE

    Article Title: Different Involvement of Promoter Methylation in the Expression of Organic Cation/Carnitine Transporter 2 (OCTN2) in Cancer Cell Lines

    doi: 10.1371/journal.pone.0076474

    Figure Lengend Snippet: Expression of OCTN2 in cancer cell lines. (A), analysis of mRNA levels of OCTN2 in cancer cell lines. Total RNA in cancer cells was purified by TRIzol reagent for cDNA synthesis and the mRNA levels of OCTN2 were analyzed by Quantitative RT-PCR. (B), analysis of protein levels of OCTN2 in cancer cells. Total proteins were extracted and protein levels of OCTN2 were analyzed by western blotting. The relative expression of mRNA and protein was normalized to β-actin. The expression of HepG2 was set as 1. All of mRNA and protein analysis were performed three times. Significant difference from HepG2 or LS174T cell is denoted with asterisk (*, p

    Article Snippet: RNA purification, cDNA Synthesis and Quantitative Real-time PCR Total cellular RNA was isolated from the treated cells using TRIzol reagent. cDNA was synthesized from 1 μg of total RNA using the first cDNA synthesis kit (Takara, Dalian).

    Techniques: Expressing, Purification, Quantitative RT-PCR, Western Blot

    Messenger RNA expression of TNC receptors of MCF-7 cells after TGF-β and TNC treatments. ( a ) mRNA was extracted from cells co-stimulated with TNC (10 μg/ml) and TGF-β1 (5 ng/ml) for synthesis of cDNA and performance of real-time PCR determining the cycle threshold ( C t ) values for each subunit. Differences in C t values of TNC receptor mRNAs from that of GAPDH mRNA in TGF-β1/TNC-treated cells are shown using qPCR. C t values of α7/8/9 and β3 integrin subunits were very high, indicating only limited expression. Calibration curves were obtained using each target gene in the samples. ( b ) Changes of receptor mRNA expression for αv/2 and β1/6 after treatment with TGF-β1 and/or TNC. The results were normalized to GAPDH expression. Integrin αv showed significant increase after TGF-β1/TNC treatment and β6 was dramatically increased after TGF-β1 addition, with or without TNC (mean±s.d.; * P

    Journal: Oncogenesis

    Article Title: Binding of ?v?1 and ?v?6 integrins to tenascin-C induces epithelial-mesenchymal transition-like change of breast cancer cells

    doi: 10.1038/oncsis.2013.27

    Figure Lengend Snippet: Messenger RNA expression of TNC receptors of MCF-7 cells after TGF-β and TNC treatments. ( a ) mRNA was extracted from cells co-stimulated with TNC (10 μg/ml) and TGF-β1 (5 ng/ml) for synthesis of cDNA and performance of real-time PCR determining the cycle threshold ( C t ) values for each subunit. Differences in C t values of TNC receptor mRNAs from that of GAPDH mRNA in TGF-β1/TNC-treated cells are shown using qPCR. C t values of α7/8/9 and β3 integrin subunits were very high, indicating only limited expression. Calibration curves were obtained using each target gene in the samples. ( b ) Changes of receptor mRNA expression for αv/2 and β1/6 after treatment with TGF-β1 and/or TNC. The results were normalized to GAPDH expression. Integrin αv showed significant increase after TGF-β1/TNC treatment and β6 was dramatically increased after TGF-β1 addition, with or without TNC (mean±s.d.; * P

    Article Snippet: Total RNA (1 μg) was reverse-transcribed using a cDNA synthesis kit (Roche Diagnostics) with anchored-oligo (dT)18 primers.

    Techniques: RNA Expression, Real-time Polymerase Chain Reaction, Expressing

    Flow chart for comprehensive NF1 mutation detection. Point mutations identified by DNA sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using cDNA sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.

    Journal: Medicine

    Article Title: Clinical and Molecular Characterization of NF1 Patients

    doi: 10.1097/MD.0000000000003043

    Figure Lengend Snippet: Flow chart for comprehensive NF1 mutation detection. Point mutations identified by DNA sequencing with specific primers (step 1) represented 68.8% of the NF1 mutations. Frameshift and nonsense mutations were identified in 31.3% and 18.8% of NF1 patients, respectively. In addition, missense and splice-site mutations were confirmed using cDNA sequencing (step 2) and were observed in 12.5% and 6.3% of NF1 patients, respectively. In the case of a negative result using DNA sequencing, an analysis of NF1 complete and large partial deletions was performed using multiplex ligation-dependent probe amplification (MLPA) (step 3) and occurred in 25.0% of NF1 patients. This comprehensive mutation screening procedure enabled us to identify an NF1 mutation in 93.8% of the NF1 patients in our study. NF1 = neurofibromatosis type 1.

    Article Snippet: To prevent illegitimate splicing, blood samples were processed after venipuncture with a maximum delay of 4 h and samples were not stored at 4°C., Reverse transcription was performed using 500 ng of total RNA isolated and random hexamers with a First-Strand complementary DNA (cDNA) Synthesis Kit for RT-PCR (AMV) (Roche Applied Science, Indianapolis, IN).

    Techniques: Flow Cytometry, Mutagenesis, DNA Sequencing, Sequencing, Multiplex Assay, Ligation, Amplification, Multiplex Ligation-dependent Probe Amplification

    The nucleotide and deduced amino acid sequences of Fus p 4.0101 (GenBank accession no. KF151224). Numbers to the right are the positions of the nucleotides and the deduced residues of the sequences. The stop codon TAA is denoted with an asterisk (*). The potential N-glycosylation site is indicated in bold letters and boxed. Eight conserved amino acid residues that are involved in the catalysis and the substrate binding of transaldolases are shaded and in bold letters. Nucleotide sequences in grey correspond to primer sequences synthesized for PCR experiments in the cDNA cloning of Fus p 4.0101 as described in the Materials and Methods. The sequences corresponding to primers Fu-TAase-f and Fu-TAase-r used in the preparation of rFus p 4.0101 are boxed.

    Journal: PLoS ONE

    Article Title: The Transaldolase, a Novel Allergen of Fusarium proliferatum, Demonstrates IgE Cross-Reactivity with Its Human Analogue

    doi: 10.1371/journal.pone.0103488

    Figure Lengend Snippet: The nucleotide and deduced amino acid sequences of Fus p 4.0101 (GenBank accession no. KF151224). Numbers to the right are the positions of the nucleotides and the deduced residues of the sequences. The stop codon TAA is denoted with an asterisk (*). The potential N-glycosylation site is indicated in bold letters and boxed. Eight conserved amino acid residues that are involved in the catalysis and the substrate binding of transaldolases are shaded and in bold letters. Nucleotide sequences in grey correspond to primer sequences synthesized for PCR experiments in the cDNA cloning of Fus p 4.0101 as described in the Materials and Methods. The sequences corresponding to primers Fu-TAase-f and Fu-TAase-r used in the preparation of rFus p 4.0101 are boxed.

    Article Snippet: cDNA Cloning The cDNA encoding the F. proliferatum transaldolase was isolated with polymerase chain reactions (PCR) using an AffinityScript Multiple Temperature cDNA Synthesis kit (Stratagene, La Jolla, Calif., USA) as previously described , .

    Techniques: Binding Assay, Synthesized, Polymerase Chain Reaction, Clone Assay

    Identification of Oplr16 as a pluripotent lncRNA. ( A ) Identification of Oplr16 as a pluripotency-associated lncRNA by integrating RNA-seq and CRIST-seq. RNA-seq was used to identify RNAs that are differentially expressed between fibroblasts and iPSCs collected in the process of reprogramming. In CRIST-seq, the Oct4 promoter was targeted by the expression of the Cas9 Oct4 -gRNA. LncRNAs interacting with Oct4 promoter were reverse transcribed in situ into cDNA with biotin-dCTP. After immunoprecipitation, biotin-cDNAs were purified by streptavidin beads for Illumina sequencing. Both sets of databases were integrated to identify the pluripotency-associated lncRNA Oplr16 . ( B ) Reactivation of Oplr16 in reprogramming. Fibroblasts were transfected with a Oct4-Sox2-Kilf4-c-Myc (OSKM) lentivirus. Cells were collected at various stages of reprogramming and expression of Oplr16 was measured by qPCR. E14: mouse embryonic pluripotent stem cell line used as a positive control; iPSC: induced pluripotent stem cells; non-iPSC: un-reprogrammed cells that express four OSKM factors, but fail to complete reprogramming; FBC: fibroblasts. β-Actin was used as the PCR control. ** P

    Journal: Nucleic Acids Research

    Article Title: Oplr16 serves as a novel chromatin factor to control stem cell fate by modulating pluripotency-specific chromosomal looping and TET2-mediated DNA demethylation

    doi: 10.1093/nar/gkaa097

    Figure Lengend Snippet: Identification of Oplr16 as a pluripotent lncRNA. ( A ) Identification of Oplr16 as a pluripotency-associated lncRNA by integrating RNA-seq and CRIST-seq. RNA-seq was used to identify RNAs that are differentially expressed between fibroblasts and iPSCs collected in the process of reprogramming. In CRIST-seq, the Oct4 promoter was targeted by the expression of the Cas9 Oct4 -gRNA. LncRNAs interacting with Oct4 promoter were reverse transcribed in situ into cDNA with biotin-dCTP. After immunoprecipitation, biotin-cDNAs were purified by streptavidin beads for Illumina sequencing. Both sets of databases were integrated to identify the pluripotency-associated lncRNA Oplr16 . ( B ) Reactivation of Oplr16 in reprogramming. Fibroblasts were transfected with a Oct4-Sox2-Kilf4-c-Myc (OSKM) lentivirus. Cells were collected at various stages of reprogramming and expression of Oplr16 was measured by qPCR. E14: mouse embryonic pluripotent stem cell line used as a positive control; iPSC: induced pluripotent stem cells; non-iPSC: un-reprogrammed cells that express four OSKM factors, but fail to complete reprogramming; FBC: fibroblasts. β-Actin was used as the PCR control. ** P

    Article Snippet: The second strand cDNA was synthesized by using Stratagene cDNA Synthesis kit (Agilent Technologies, CA, USA) for Illumina lnc-cDNA sequencing.

    Techniques: RNA Sequencing Assay, Expressing, In Situ, Immunoprecipitation, Purification, Sequencing, Transfection, Real-time Polymerase Chain Reaction, Positive Control, Polymerase Chain Reaction

    Oplr16 binds to the Oct4 promoter. ( A ) Schematic diagram of the RNA reverse transcription-associated trap sequencing (RAT-seq) assay. Oplr16 lncRNA was in situ reverse transcribed using three Oplr16 -specific complementary primers with biotin-dCTP. The random primers were used as the negative control (RAT-CT). After nuclear lysis, the bitoin- Oplr16 cDNA chromatin complex was isolated by streptavidin beads and the Oplr16 -interacting target DNAs were isolated for Illumina library sequencing. ( B ) The IGV analysis of Oplr16 binding signals in the Oct4 locus. 5′-Enh, 3′-Enh: the Oct4 5′- and 3′-enhancers; pOct4: Oct4 promoter; E1-E5: Oct4 exons. ( C ) Quantitative PCR mapping of Oplr16 binding in the Oct4 locus. The RAT pulldown complex was used to map the Oplr16 binding. 5′-CT, 3′-CT: the RAT control sites in the Oct4 locus. Note the enrichment of the Oplr16 binding signals in the promoter region (Prot-1 and Prot-2) ( N = 9, ** P

    Journal: Nucleic Acids Research

    Article Title: Oplr16 serves as a novel chromatin factor to control stem cell fate by modulating pluripotency-specific chromosomal looping and TET2-mediated DNA demethylation

    doi: 10.1093/nar/gkaa097

    Figure Lengend Snippet: Oplr16 binds to the Oct4 promoter. ( A ) Schematic diagram of the RNA reverse transcription-associated trap sequencing (RAT-seq) assay. Oplr16 lncRNA was in situ reverse transcribed using three Oplr16 -specific complementary primers with biotin-dCTP. The random primers were used as the negative control (RAT-CT). After nuclear lysis, the bitoin- Oplr16 cDNA chromatin complex was isolated by streptavidin beads and the Oplr16 -interacting target DNAs were isolated for Illumina library sequencing. ( B ) The IGV analysis of Oplr16 binding signals in the Oct4 locus. 5′-Enh, 3′-Enh: the Oct4 5′- and 3′-enhancers; pOct4: Oct4 promoter; E1-E5: Oct4 exons. ( C ) Quantitative PCR mapping of Oplr16 binding in the Oct4 locus. The RAT pulldown complex was used to map the Oplr16 binding. 5′-CT, 3′-CT: the RAT control sites in the Oct4 locus. Note the enrichment of the Oplr16 binding signals in the promoter region (Prot-1 and Prot-2) ( N = 9, ** P

    Article Snippet: The second strand cDNA was synthesized by using Stratagene cDNA Synthesis kit (Agilent Technologies, CA, USA) for Illumina lnc-cDNA sequencing.

    Techniques: Sequencing, In Situ, Negative Control, Lysis, Isolation, Binding Assay, Real-time Polymerase Chain Reaction