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  • 91
    Thermo Fisher cdk6 complementary dna cdna
    Cdk6 Complementary Dna Cdna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Bio-Rad complementary dna cdna preparation
    Complementary Dna Cdna Preparation, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 94 article reviews
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    88
    Toyobo complementary dna cdna preparation
    Complementary Dna Cdna Preparation, supplied by Toyobo, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Roche prepared complementary dna
    Prepared Complementary Dna, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Roche cdna preparation
    Differential expression of ALCAM in pancreatic cancer cells and PSCs. (A) Reverse transcription-quantitative polymerase chain reaction analysis of ALCAM mRNA levels in the pancreatic cancer cell lines Panc-1 and T3M4, as well as in PSCs. RNA input was normalized against the average expression of hypoxanthine-guanine phosphoribosyltransferase and cylophilin B, and presented as the copy number/ µ l <t>cDNA.</t> Immunoblotting analysis was performed to detect ALCAM protein expression in Panc-1 and T3M4 cells, and also in PSCs. (B) Equal loading of the protein samples was confirmed using an ERK-2 antibody. ALCAM, activated leukocyte cell adhesion molecule; PSCs, pancreatic stellate cells; ERK-2, extracellular signal-regulated kinase-2.
    Cdna Preparation, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 357 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bioarray complementary dna preparation
    Differential expression of ALCAM in pancreatic cancer cells and PSCs. (A) Reverse transcription-quantitative polymerase chain reaction analysis of ALCAM mRNA levels in the pancreatic cancer cell lines Panc-1 and T3M4, as well as in PSCs. RNA input was normalized against the average expression of hypoxanthine-guanine phosphoribosyltransferase and cylophilin B, and presented as the copy number/ µ l <t>cDNA.</t> Immunoblotting analysis was performed to detect ALCAM protein expression in Panc-1 and T3M4 cells, and also in PSCs. (B) Equal loading of the protein samples was confirmed using an ERK-2 antibody. ALCAM, activated leukocyte cell adhesion molecule; PSCs, pancreatic stellate cells; ERK-2, extracellular signal-regulated kinase-2.
    Complementary Dna Preparation, supplied by Bioarray, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    TaKaRa complementary dna cdna
    Construction of pFastBac vectors. (A) Toxoplasma gondii IMC gene was PCR-amplified from <t>cDNA</t> synthesized using a Prime Script 1st Strain CDNA Synthesis Kit using total RNA extracted from T . gondii RH. M denotes <t>DNA</t> marker and lane 1 indicates amplified PCR product. (B) Influenza M1 gene was PCR amplified from total RNA extracted from influenza virus (A/PR/8/34). M denotes DNA marker and lane 2 indicates amplified PCR product. (C and D) Toxoplasma gondii IMC gene and influenza M1 gene were cloned in to pFastBac with EcoRI / XhoI and EcoRI / XhoI enzymes, respectively, resulting in T . gondii IMC plasmid (C) and M1 plasmid (D).
    Complementary Dna Cdna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 6487 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Genecopoeia maf complementary dna
    Construction of pFastBac vectors. (A) Toxoplasma gondii IMC gene was PCR-amplified from <t>cDNA</t> synthesized using a Prime Script 1st Strain CDNA Synthesis Kit using total RNA extracted from T . gondii RH. M denotes <t>DNA</t> marker and lane 1 indicates amplified PCR product. (B) Influenza M1 gene was PCR amplified from total RNA extracted from influenza virus (A/PR/8/34). M denotes DNA marker and lane 2 indicates amplified PCR product. (C and D) Toxoplasma gondii IMC gene and influenza M1 gene were cloned in to pFastBac with EcoRI / XhoI and EcoRI / XhoI enzymes, respectively, resulting in T . gondii IMC plasmid (C) and M1 plasmid (D).
    Maf Complementary Dna, supplied by Genecopoeia, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 90 stars, based on 7 article reviews
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    90
    Addgene inc complementary dna cdna
    Construction of pFastBac vectors. (A) Toxoplasma gondii IMC gene was PCR-amplified from <t>cDNA</t> synthesized using a Prime Script 1st Strain CDNA Synthesis Kit using total RNA extracted from T . gondii RH. M denotes <t>DNA</t> marker and lane 1 indicates amplified PCR product. (B) Influenza M1 gene was PCR amplified from total RNA extracted from influenza virus (A/PR/8/34). M denotes DNA marker and lane 2 indicates amplified PCR product. (C and D) Toxoplasma gondii IMC gene and influenza M1 gene were cloned in to pFastBac with EcoRI / XhoI and EcoRI / XhoI enzymes, respectively, resulting in T . gondii IMC plasmid (C) and M1 plasmid (D).
    Complementary Dna Cdna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Nugen complementary dna cdna
    Construction of pFastBac vectors. (A) Toxoplasma gondii IMC gene was PCR-amplified from <t>cDNA</t> synthesized using a Prime Script 1st Strain CDNA Synthesis Kit using total RNA extracted from T . gondii RH. M denotes <t>DNA</t> marker and lane 1 indicates amplified PCR product. (B) Influenza M1 gene was PCR amplified from total RNA extracted from influenza virus (A/PR/8/34). M denotes DNA marker and lane 2 indicates amplified PCR product. (C and D) Toxoplasma gondii IMC gene and influenza M1 gene were cloned in to pFastBac with EcoRI / XhoI and EcoRI / XhoI enzymes, respectively, resulting in T . gondii IMC plasmid (C) and M1 plasmid (D).
    Complementary Dna Cdna, supplied by Nugen, used in various techniques. Bioz Stars score: 93/100, based on 149 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher complementary dna cdna
    FAXDC2 expedites megakaryopoiesis in murine bone marrow cells. ( a ) c-kit-positive progenitor cells isolated from primary murine bone marrow cells were transduced with control lentiviral vector (Ctrl) or FAXDC2-overexpressing (FAXDC2) vector to undergo megakaryocytic differentiation with thrombopoietin (TPO) for 6 days. The resultant cells were stained with phycoerythrin (PE)-CY7-conjugated anti-CD41 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( b ) The resultant cells were stained with PE-conjugated anti CD42 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( c ) The resultant cells were stained with anti-CD41-PE-CY7 and DAPI. The CD41 + cells were gated for analysis of <t>DNA</t> content. The DNA content that is > 4N is represented (left panel). Bar graph (right panel) was the statistics of left panel. ( d ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for Wright-Giemsa stains. The stained cells were photographed under microscopy at the bright view of the microscope (magnification × 20 or magnification × 40). Scale Bar: 100 μm. ( e ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for quantitative real-time <t>PCR</t> (RT-PCR) at mRNA level. The expression of RUNX1 was measured. * P
    Complementary Dna Cdna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 27072 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Transgenomic complementary dna cdna
    FAXDC2 expedites megakaryopoiesis in murine bone marrow cells. ( a ) c-kit-positive progenitor cells isolated from primary murine bone marrow cells were transduced with control lentiviral vector (Ctrl) or FAXDC2-overexpressing (FAXDC2) vector to undergo megakaryocytic differentiation with thrombopoietin (TPO) for 6 days. The resultant cells were stained with phycoerythrin (PE)-CY7-conjugated anti-CD41 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( b ) The resultant cells were stained with PE-conjugated anti CD42 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( c ) The resultant cells were stained with anti-CD41-PE-CY7 and DAPI. The CD41 + cells were gated for analysis of <t>DNA</t> content. The DNA content that is > 4N is represented (left panel). Bar graph (right panel) was the statistics of left panel. ( d ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for Wright-Giemsa stains. The stained cells were photographed under microscopy at the bright view of the microscope (magnification × 20 or magnification × 40). Scale Bar: 100 μm. ( e ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for quantitative real-time <t>PCR</t> (RT-PCR) at mRNA level. The expression of RUNX1 was measured. * P
    Complementary Dna Cdna, supplied by Transgenomic, used in various techniques. Bioz Stars score: 92/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Addgene inc full length myap complementary dna cdna
    FAXDC2 expedites megakaryopoiesis in murine bone marrow cells. ( a ) c-kit-positive progenitor cells isolated from primary murine bone marrow cells were transduced with control lentiviral vector (Ctrl) or FAXDC2-overexpressing (FAXDC2) vector to undergo megakaryocytic differentiation with thrombopoietin (TPO) for 6 days. The resultant cells were stained with phycoerythrin (PE)-CY7-conjugated anti-CD41 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( b ) The resultant cells were stained with PE-conjugated anti CD42 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( c ) The resultant cells were stained with anti-CD41-PE-CY7 and DAPI. The CD41 + cells were gated for analysis of <t>DNA</t> content. The DNA content that is > 4N is represented (left panel). Bar graph (right panel) was the statistics of left panel. ( d ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for Wright-Giemsa stains. The stained cells were photographed under microscopy at the bright view of the microscope (magnification × 20 or magnification × 40). Scale Bar: 100 μm. ( e ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for quantitative real-time <t>PCR</t> (RT-PCR) at mRNA level. The expression of RUNX1 was measured. * P
    Full Length Myap Complementary Dna Cdna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 85 stars, based on 5 article reviews
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    88
    TaKaRa complementary dna oligos
    FAXDC2 expedites megakaryopoiesis in murine bone marrow cells. ( a ) c-kit-positive progenitor cells isolated from primary murine bone marrow cells were transduced with control lentiviral vector (Ctrl) or FAXDC2-overexpressing (FAXDC2) vector to undergo megakaryocytic differentiation with thrombopoietin (TPO) for 6 days. The resultant cells were stained with phycoerythrin (PE)-CY7-conjugated anti-CD41 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( b ) The resultant cells were stained with PE-conjugated anti CD42 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( c ) The resultant cells were stained with anti-CD41-PE-CY7 and DAPI. The CD41 + cells were gated for analysis of <t>DNA</t> content. The DNA content that is > 4N is represented (left panel). Bar graph (right panel) was the statistics of left panel. ( d ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for Wright-Giemsa stains. The stained cells were photographed under microscopy at the bright view of the microscope (magnification × 20 or magnification × 40). Scale Bar: 100 μm. ( e ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for quantitative real-time <t>PCR</t> (RT-PCR) at mRNA level. The expression of RUNX1 was measured. * P
    Complementary Dna Oligos, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher human mcm7 complementary dna cdna
    Pb/MCM7i13 transgenic construct. (A) Pb, prostate-specific rat probasin promoter ARR 2 PB; black box, HA tag; white box, Xpress tag; gray boxes, human <t>MCM7</t> coding sequence; red and blue boxes, miR-93/106b and miR-25 genes; yellow box, SV40 poly(A) (SV40 pA); orange box, probe used for Southern analysis of transgenic lines; green arrows, primers used for PCR analysis of transgenic lines and for genotyping. (B) Top: Southern blot analysis. Genomic <t>DNA</t> extracted from tails was digested with Bam HI and hybridized with a probe complementary to the SV40 poly(A) region as shown in (A). The Bam HI–digested Pb/MCM7i13 plasmid was used as positive control (C+). Bottom: PCR analysis. A forward primer complementary to the probasin promoter and a reverse primer complementary to the tag were used, as shown in (A). Empty Pb and Pb/MCM7i13 plasmids were used as negative (C−) and positive (C+) controls. L, 100-bp ladder. Three representative positive (P) and negative (N) mice are shown. (C) Abundance of the transgenic construct measured by real-time PCR in nine different lines. A primer pair specific for human MCM7 coding sequence was used. LE, ME, HE: low, medium, and high expressors, respectively. Human MCM7 .
    Human Mcm7 Complementary Dna Cdna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Addgene inc complementary dna cdna fragment
    Pb/MCM7i13 transgenic construct. (A) Pb, prostate-specific rat probasin promoter ARR 2 PB; black box, HA tag; white box, Xpress tag; gray boxes, human <t>MCM7</t> coding sequence; red and blue boxes, miR-93/106b and miR-25 genes; yellow box, SV40 poly(A) (SV40 pA); orange box, probe used for Southern analysis of transgenic lines; green arrows, primers used for PCR analysis of transgenic lines and for genotyping. (B) Top: Southern blot analysis. Genomic <t>DNA</t> extracted from tails was digested with Bam HI and hybridized with a probe complementary to the SV40 poly(A) region as shown in (A). The Bam HI–digested Pb/MCM7i13 plasmid was used as positive control (C+). Bottom: PCR analysis. A forward primer complementary to the probasin promoter and a reverse primer complementary to the tag were used, as shown in (A). Empty Pb and Pb/MCM7i13 plasmids were used as negative (C−) and positive (C+) controls. L, 100-bp ladder. Three representative positive (P) and negative (N) mice are shown. (C) Abundance of the transgenic construct measured by real-time PCR in nine different lines. A primer pair specific for human MCM7 coding sequence was used. LE, ME, HE: low, medium, and high expressors, respectively. Human MCM7 .
    Complementary Dna Cdna Fragment, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher brain complementary dna
    Pb/MCM7i13 transgenic construct. (A) Pb, prostate-specific rat probasin promoter ARR 2 PB; black box, HA tag; white box, Xpress tag; gray boxes, human <t>MCM7</t> coding sequence; red and blue boxes, miR-93/106b and miR-25 genes; yellow box, SV40 poly(A) (SV40 pA); orange box, probe used for Southern analysis of transgenic lines; green arrows, primers used for PCR analysis of transgenic lines and for genotyping. (B) Top: Southern blot analysis. Genomic <t>DNA</t> extracted from tails was digested with Bam HI and hybridized with a probe complementary to the SV40 poly(A) region as shown in (A). The Bam HI–digested Pb/MCM7i13 plasmid was used as positive control (C+). Bottom: PCR analysis. A forward primer complementary to the probasin promoter and a reverse primer complementary to the tag were used, as shown in (A). Empty Pb and Pb/MCM7i13 plasmids were used as negative (C−) and positive (C+) controls. L, 100-bp ladder. Three representative positive (P) and negative (N) mice are shown. (C) Abundance of the transgenic construct measured by real-time PCR in nine different lines. A primer pair specific for human MCM7 coding sequence was used. LE, ME, HE: low, medium, and high expressors, respectively. Human MCM7 .
    Brain Complementary Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Illumina Inc complementary dna library preparation
    Pb/MCM7i13 transgenic construct. (A) Pb, prostate-specific rat probasin promoter ARR 2 PB; black box, HA tag; white box, Xpress tag; gray boxes, human <t>MCM7</t> coding sequence; red and blue boxes, miR-93/106b and miR-25 genes; yellow box, SV40 poly(A) (SV40 pA); orange box, probe used for Southern analysis of transgenic lines; green arrows, primers used for PCR analysis of transgenic lines and for genotyping. (B) Top: Southern blot analysis. Genomic <t>DNA</t> extracted from tails was digested with Bam HI and hybridized with a probe complementary to the SV40 poly(A) region as shown in (A). The Bam HI–digested Pb/MCM7i13 plasmid was used as positive control (C+). Bottom: PCR analysis. A forward primer complementary to the probasin promoter and a reverse primer complementary to the tag were used, as shown in (A). Empty Pb and Pb/MCM7i13 plasmids were used as negative (C−) and positive (C+) controls. L, 100-bp ladder. Three representative positive (P) and negative (N) mice are shown. (C) Abundance of the transgenic construct measured by real-time PCR in nine different lines. A primer pair specific for human MCM7 coding sequence was used. LE, ME, HE: low, medium, and high expressors, respectively. Human MCM7 .
    Complementary Dna Library Preparation, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 88/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    InvivoGen complementary dna
    Pb/MCM7i13 transgenic construct. (A) Pb, prostate-specific rat probasin promoter ARR 2 PB; black box, HA tag; white box, Xpress tag; gray boxes, human <t>MCM7</t> coding sequence; red and blue boxes, miR-93/106b and miR-25 genes; yellow box, SV40 poly(A) (SV40 pA); orange box, probe used for Southern analysis of transgenic lines; green arrows, primers used for PCR analysis of transgenic lines and for genotyping. (B) Top: Southern blot analysis. Genomic <t>DNA</t> extracted from tails was digested with Bam HI and hybridized with a probe complementary to the SV40 poly(A) region as shown in (A). The Bam HI–digested Pb/MCM7i13 plasmid was used as positive control (C+). Bottom: PCR analysis. A forward primer complementary to the probasin promoter and a reverse primer complementary to the tag were used, as shown in (A). Empty Pb and Pb/MCM7i13 plasmids were used as negative (C−) and positive (C+) controls. L, 100-bp ladder. Three representative positive (P) and negative (N) mice are shown. (C) Abundance of the transgenic construct measured by real-time PCR in nine different lines. A primer pair specific for human MCM7 coding sequence was used. LE, ME, HE: low, medium, and high expressors, respectively. Human MCM7 .
    Complementary Dna, supplied by InvivoGen, used in various techniques. Bioz Stars score: 92/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher human crbn complementary dna
    Pb/MCM7i13 transgenic construct. (A) Pb, prostate-specific rat probasin promoter ARR 2 PB; black box, HA tag; white box, Xpress tag; gray boxes, human <t>MCM7</t> coding sequence; red and blue boxes, miR-93/106b and miR-25 genes; yellow box, SV40 poly(A) (SV40 pA); orange box, probe used for Southern analysis of transgenic lines; green arrows, primers used for PCR analysis of transgenic lines and for genotyping. (B) Top: Southern blot analysis. Genomic <t>DNA</t> extracted from tails was digested with Bam HI and hybridized with a probe complementary to the SV40 poly(A) region as shown in (A). The Bam HI–digested Pb/MCM7i13 plasmid was used as positive control (C+). Bottom: PCR analysis. A forward primer complementary to the probasin promoter and a reverse primer complementary to the tag were used, as shown in (A). Empty Pb and Pb/MCM7i13 plasmids were used as negative (C−) and positive (C+) controls. L, 100-bp ladder. Three representative positive (P) and negative (N) mice are shown. (C) Abundance of the transgenic construct measured by real-time PCR in nine different lines. A primer pair specific for human MCM7 coding sequence was used. LE, ME, HE: low, medium, and high expressors, respectively. Human MCM7 .
    Human Crbn Complementary Dna, supplied by Thermo Fisher, 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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    Image Search Results


    Differential expression of ALCAM in pancreatic cancer cells and PSCs. (A) Reverse transcription-quantitative polymerase chain reaction analysis of ALCAM mRNA levels in the pancreatic cancer cell lines Panc-1 and T3M4, as well as in PSCs. RNA input was normalized against the average expression of hypoxanthine-guanine phosphoribosyltransferase and cylophilin B, and presented as the copy number/ µ l cDNA. Immunoblotting analysis was performed to detect ALCAM protein expression in Panc-1 and T3M4 cells, and also in PSCs. (B) Equal loading of the protein samples was confirmed using an ERK-2 antibody. ALCAM, activated leukocyte cell adhesion molecule; PSCs, pancreatic stellate cells; ERK-2, extracellular signal-regulated kinase-2.

    Journal: Molecular Medicine Reports

    Article Title: Activated leukocyte cell adhesion molecule regulates the interaction between pancreatic cancer cells and stellate cells

    doi: 10.3892/mmr.2016.5681

    Figure Lengend Snippet: Differential expression of ALCAM in pancreatic cancer cells and PSCs. (A) Reverse transcription-quantitative polymerase chain reaction analysis of ALCAM mRNA levels in the pancreatic cancer cell lines Panc-1 and T3M4, as well as in PSCs. RNA input was normalized against the average expression of hypoxanthine-guanine phosphoribosyltransferase and cylophilin B, and presented as the copy number/ µ l cDNA. Immunoblotting analysis was performed to detect ALCAM protein expression in Panc-1 and T3M4 cells, and also in PSCs. (B) Equal loading of the protein samples was confirmed using an ERK-2 antibody. ALCAM, activated leukocyte cell adhesion molecule; PSCs, pancreatic stellate cells; ERK-2, extracellular signal-regulated kinase-2.

    Article Snippet: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) All reagents and equipment for mRNA and cDNA preparation were purchased from Roche (Mannheim, Germany).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Construction of pFastBac vectors. (A) Toxoplasma gondii IMC gene was PCR-amplified from cDNA synthesized using a Prime Script 1st Strain CDNA Synthesis Kit using total RNA extracted from T . gondii RH. M denotes DNA marker and lane 1 indicates amplified PCR product. (B) Influenza M1 gene was PCR amplified from total RNA extracted from influenza virus (A/PR/8/34). M denotes DNA marker and lane 2 indicates amplified PCR product. (C and D) Toxoplasma gondii IMC gene and influenza M1 gene were cloned in to pFastBac with EcoRI / XhoI and EcoRI / XhoI enzymes, respectively, resulting in T . gondii IMC plasmid (C) and M1 plasmid (D).

    Journal: PLoS ONE

    Article Title: Virus-Like Nanoparticle Vaccine Confers Protection against Toxoplasma gondii

    doi: 10.1371/journal.pone.0161231

    Figure Lengend Snippet: Construction of pFastBac vectors. (A) Toxoplasma gondii IMC gene was PCR-amplified from cDNA synthesized using a Prime Script 1st Strain CDNA Synthesis Kit using total RNA extracted from T . gondii RH. M denotes DNA marker and lane 1 indicates amplified PCR product. (B) Influenza M1 gene was PCR amplified from total RNA extracted from influenza virus (A/PR/8/34). M denotes DNA marker and lane 2 indicates amplified PCR product. (C and D) Toxoplasma gondii IMC gene and influenza M1 gene were cloned in to pFastBac with EcoRI / XhoI and EcoRI / XhoI enzymes, respectively, resulting in T . gondii IMC plasmid (C) and M1 plasmid (D).

    Article Snippet: Complementary DNA (cDNA) was synthesized using a Prime Script 1st Strain CDNA Synthesis Kit (Takara, Japan).

    Techniques: Polymerase Chain Reaction, Amplification, Synthesized, Marker, Clone Assay, Plasmid Preparation

    FAXDC2 expedites megakaryopoiesis in murine bone marrow cells. ( a ) c-kit-positive progenitor cells isolated from primary murine bone marrow cells were transduced with control lentiviral vector (Ctrl) or FAXDC2-overexpressing (FAXDC2) vector to undergo megakaryocytic differentiation with thrombopoietin (TPO) for 6 days. The resultant cells were stained with phycoerythrin (PE)-CY7-conjugated anti-CD41 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( b ) The resultant cells were stained with PE-conjugated anti CD42 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( c ) The resultant cells were stained with anti-CD41-PE-CY7 and DAPI. The CD41 + cells were gated for analysis of DNA content. The DNA content that is > 4N is represented (left panel). Bar graph (right panel) was the statistics of left panel. ( d ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for Wright-Giemsa stains. The stained cells were photographed under microscopy at the bright view of the microscope (magnification × 20 or magnification × 40). Scale Bar: 100 μm. ( e ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for quantitative real-time PCR (RT-PCR) at mRNA level. The expression of RUNX1 was measured. * P

    Journal: Blood Cancer Journal

    Article Title: Novel function of FAXDC2 in megakaryopoiesis

    doi: 10.1038/bcj.2016.87

    Figure Lengend Snippet: FAXDC2 expedites megakaryopoiesis in murine bone marrow cells. ( a ) c-kit-positive progenitor cells isolated from primary murine bone marrow cells were transduced with control lentiviral vector (Ctrl) or FAXDC2-overexpressing (FAXDC2) vector to undergo megakaryocytic differentiation with thrombopoietin (TPO) for 6 days. The resultant cells were stained with phycoerythrin (PE)-CY7-conjugated anti-CD41 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( b ) The resultant cells were stained with PE-conjugated anti CD42 antibody and analyzed by flow cytometry (left panel). Bar graph (right panel) was the statistics of left panel. ( c ) The resultant cells were stained with anti-CD41-PE-CY7 and DAPI. The CD41 + cells were gated for analysis of DNA content. The DNA content that is > 4N is represented (left panel). Bar graph (right panel) was the statistics of left panel. ( d ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for Wright-Giemsa stains. The stained cells were photographed under microscopy at the bright view of the microscope (magnification × 20 or magnification × 40). Scale Bar: 100 μm. ( e ) The control (Ctrl) or FAXDC2-overexpressing (FAXDC2) cells were harvested for quantitative real-time PCR (RT-PCR) at mRNA level. The expression of RUNX1 was measured. * P

    Article Snippet: RNA preparation and quantitative real-time PCR Total RNA was extracted with Trizol and complementary DNA was prepared with reverse transcriptase by standard procedure following instructions of the manufacturer (Invitrogen, Grand Island, NY, USA).

    Techniques: Isolation, Transduction, Plasmid Preparation, Staining, Flow Cytometry, Cytometry, Microscopy, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Expressing

    Pb/MCM7i13 transgenic construct. (A) Pb, prostate-specific rat probasin promoter ARR 2 PB; black box, HA tag; white box, Xpress tag; gray boxes, human MCM7 coding sequence; red and blue boxes, miR-93/106b and miR-25 genes; yellow box, SV40 poly(A) (SV40 pA); orange box, probe used for Southern analysis of transgenic lines; green arrows, primers used for PCR analysis of transgenic lines and for genotyping. (B) Top: Southern blot analysis. Genomic DNA extracted from tails was digested with Bam HI and hybridized with a probe complementary to the SV40 poly(A) region as shown in (A). The Bam HI–digested Pb/MCM7i13 plasmid was used as positive control (C+). Bottom: PCR analysis. A forward primer complementary to the probasin promoter and a reverse primer complementary to the tag were used, as shown in (A). Empty Pb and Pb/MCM7i13 plasmids were used as negative (C−) and positive (C+) controls. L, 100-bp ladder. Three representative positive (P) and negative (N) mice are shown. (C) Abundance of the transgenic construct measured by real-time PCR in nine different lines. A primer pair specific for human MCM7 coding sequence was used. LE, ME, HE: low, medium, and high expressors, respectively. Human MCM7 .

    Journal: Science signaling

    Article Title: Identification of the miR-106b~25 MicroRNA Cluster as a Proto-Oncogenic PTEN-Targeting Intron That Cooperates with Its Host Gene MCM7 in Transformation

    doi: 10.1126/scisignal.2000594

    Figure Lengend Snippet: Pb/MCM7i13 transgenic construct. (A) Pb, prostate-specific rat probasin promoter ARR 2 PB; black box, HA tag; white box, Xpress tag; gray boxes, human MCM7 coding sequence; red and blue boxes, miR-93/106b and miR-25 genes; yellow box, SV40 poly(A) (SV40 pA); orange box, probe used for Southern analysis of transgenic lines; green arrows, primers used for PCR analysis of transgenic lines and for genotyping. (B) Top: Southern blot analysis. Genomic DNA extracted from tails was digested with Bam HI and hybridized with a probe complementary to the SV40 poly(A) region as shown in (A). The Bam HI–digested Pb/MCM7i13 plasmid was used as positive control (C+). Bottom: PCR analysis. A forward primer complementary to the probasin promoter and a reverse primer complementary to the tag were used, as shown in (A). Empty Pb and Pb/MCM7i13 plasmids were used as negative (C−) and positive (C+) controls. L, 100-bp ladder. Three representative positive (P) and negative (N) mice are shown. (C) Abundance of the transgenic construct measured by real-time PCR in nine different lines. A primer pair specific for human MCM7 coding sequence was used. LE, ME, HE: low, medium, and high expressors, respectively. Human MCM7 .

    Article Snippet: The following antibodies and reagents were used: antibodies against Hsp-90 #61041 (Becton Dickinson); PTEN #9559 (WB), pAkt (Ser473 ) #9271 (WB), #3787 (IHC), Akt #9272, pS6 #2211 (Cell Signaling); DICER #CG006 (Clonegene); MSCV-neo (Clontech); siGENOME non-targeting small interfering RNA (siRNA) #2 (si-Luc), si-miR-19a, si-miR-19b, si-miR-22, si-miR-25, si-miR-92a, si-miR-17, si-miR-20a, si-miR-93, si-miR-106b, si-miR-302a, si-miR-372, si-miR-373, si-PTEN, siGLO RISC-free control siRNA miRNA antisense inhibitor negative control #1, miR-19a antisense inhibitor, miR-22 antisense inhibitor, miR-25 antisense inhibitor, miR-93 antisense inhibitor, miR-106b antisense inhibitor, Dharmafect 1 (Dharmacon); miR-17, miR-19a, miR-20a, miR-22, miR-25, miR-93, and miR-106b 3′ digoxigenin (DIG)–labeled LNA ISH probes (Exiqon); lipofectamine 2000, Trizol reagent, deoxyribonuclease I (DNase I) amplification grade, SuperScript II reverse transcriptase, Dulbecco's modified Eagle's medium (DMEM), RPMI 1640, fetal bovine serum (FBS), keratinocyte serum-free medium, epidermal growth factor (EGF), bovine pituitary extract (BPE), antibody against Xpress Tag, pcDNA3.1/His C, geneticin (G418), hygromycin, antibody against PTEN (IHC) #51-2400 (Invitrogen); human MCM7 complementary DNA (cDNA) #MHS1011-75176 (Open Biosystems); pGL3-Control ( Firefly luciferase ), pRL-TK ( Renilla luciferase ), Dual-Luciferase reporter assay (Promega); fraction V bovine serum albumin (BSA), polybrene, insulin, antibody against actin #A3853, anti-FLAG antibody #F3165, puromycin (Sigma); QuantiTect Sybr Green PCRkit (Qiagen); antibody against MCM7 #9966 (Santa Cruz Biotechnology); QuikChange II XL Site-Directed Mutagenesis Kit, Herculase Taq polymerase (Stratagene); Ki67 antibody #VP-K451 (Vector Laboratories).

    Techniques: Transgenic Assay, Construct, Sequencing, Polymerase Chain Reaction, Southern Blot, Plasmid Preparation, Positive Control, Mouse Assay, Real-time Polymerase Chain Reaction