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  • 99
    Millipore complementary dna templates
    Complementary Dna Templates, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher complementary dna cdna templates
    Complementary Dna Cdna Templates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher complementary dna cdna template
    Expression levels of <t>XYLP</t> mRNA in different human tissues. Multiple tissue <t>cDNA</t> panels (Clontech) were used to perform real time PCR analyses. Levels of XYLP ( open bars ) and FAM20B ( closed bars ) mRNAs were normalized to those of glyceraldehyde-3-phosphate
    Complementary Dna Cdna Template, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Bio-Rad complementary dna cdna template
    In vivo deletion of Tlk1 and Tlk2 in mouse mammary fibroblasts increases proliferation of adjacent ductal epithelium (A, B) Schematic overview of the targeting vector used to generate the Tlk1 and Tlk2 conditional knock-out mice. Line 1: partial genomic locus of mouse Tlk1 or Tlk2 gene with exons represented as dark gray boxes and Southern probe as light gray bar. Line 2: targeting vector with exons 9 and 10 in Tlk1 and exon 11 in Tlk2 flanked by loxP elements (black triangles), the LacZ reporter cassette and neomycin (Neo) resistance cassette flanked by FRT sites (white triangles). Line 3: targeted genomic locus (Neo). Line 4: Neo cassette was removed by Actin-Flpe mediated recombination ( Flox (F) ). Black arrows P1–P3 indicate location of genotyping primers. Diagram not to scale. (C, D) Southern blot (left panels) of ES cells confirming targeting of Tlk1 or Tlk2 genomic loci respectively. <t>PCR</t> genotyping of genomic DNA (right panels) from WT ( Tlk1 +/+ or Tlk2 +/+ ) heterozygous ( Tlk1 +/F or Tlk2 +/F ) and homozygous ( Tlk1 F/F or Tlk2 F/F ) mice with primers P1/P2 following deletion of Neo cassette. (E) Left: bright field images of whole mount inguinal mammary glands from 10-week-old Tlk1 F/F ;Tlk2 F/F (control) and Fsp-cre;Tlk1 F/F ;Tlk2 F/F (Tlk1 Δ/Δ ;Tlk2 Δ/Δ ) mice stained with carmine red (bar = 2000 μm). Boxes indicate area of magnification in adjacent panel (bar = 100 μm). Middle: H E stained sections of opposite inguinal gland (bar = 50 μm). Right: representative images of sections of inguinal mammary glands immuno-stained for the proliferation marker Ki67 (brown) and counterstained with hematoxylin (blue) (bar = 50 μm). Boxes indicate area of magnification in adjacent panel (bar = 20 μm). (F) Percent Ki67 positive epithelial cells per 20X field (dots) of tissue sections described in (E). Red bar, median; black bars, interquartile range; pooled counts from 6 control mice or 5 Fsp-cre mice. P value: Mann-Whitney test of medians. (G) Left: Representative images of inguinal mammary gland sections immuno-stained for smooth muscle actin (SMA; brown) and counterstained with hematoxylin (blue) (bar = 50 μm). Insets represent consecutive sections immuno-stained with Ki67 (brown) and counterstained with hematoxylin (blue). Right: Representative images of inguinal mammary gland sections immuno-stained for smooth muscle actin (SMA; red) and Ki67 (brown); counterstained with hematoxylin (blue) (bar = 50 μm). (H) PCR genotyping of mammary fibroblasts derived from Tlk1 F/F and Tlk2 F/F mice with or without Fsp-cre to detect the Flox allele ( Tlk F ; P1/P2 primers), the deletion product formed after Fsp-cre -mediated recombination ( Tlk Δ ; P1/P3 primers) and Fsp-cre. (I) qRT-PCR for Tlk1 or Tlk2 on <t>cDNA</t> derived from fibroblasts isolated from Tlk1 F/F ;Tlk2 F/F mammary glands of control (−) and Fsp-cre positive (+) mice. Expression was normalized to Gapdh. (J) Mammary gland wholemounts from Fsp-cre;Rosa loxP/+ and Rosa loxP/+ (left, inset) mice stained for β-galactosidase. Higher (10×) magnification of a wholemount gland (top-right) and a cross section stained for β-galactosidase and counterstained with nuclear fast red (bottom-right). Str, stromal fibroblasts; lu, lumen; epi, epithelial cells. (K) qRT-PCR for Tlk1 expression (left) on cDNA derived from Tlk1 F/F mammary fibroblasts (MMFs) or mammary epithelial cells (MECs) lacking (−) or containing (+) the Fsp-cre transgene. qRT-PCR for Tlk2 expression (right) on cDNA derived from Tlk2 F/F mammary fibroblasts (MMFs) or mammary epithelial cells (MECs) lacking (−) or containing (+) the Fsp-cre transgene. Expression was normalized to Gapdh. .
    Complementary Dna Cdna Template, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 88/100, based on 577 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Bio-Rad complementary dna cdna templates
    In vivo deletion of Tlk1 and Tlk2 in mouse mammary fibroblasts increases proliferation of adjacent ductal epithelium (A, B) Schematic overview of the targeting vector used to generate the Tlk1 and Tlk2 conditional knock-out mice. Line 1: partial genomic locus of mouse Tlk1 or Tlk2 gene with exons represented as dark gray boxes and Southern probe as light gray bar. Line 2: targeting vector with exons 9 and 10 in Tlk1 and exon 11 in Tlk2 flanked by loxP elements (black triangles), the LacZ reporter cassette and neomycin (Neo) resistance cassette flanked by FRT sites (white triangles). Line 3: targeted genomic locus (Neo). Line 4: Neo cassette was removed by Actin-Flpe mediated recombination ( Flox (F) ). Black arrows P1–P3 indicate location of genotyping primers. Diagram not to scale. (C, D) Southern blot (left panels) of ES cells confirming targeting of Tlk1 or Tlk2 genomic loci respectively. <t>PCR</t> genotyping of genomic DNA (right panels) from WT ( Tlk1 +/+ or Tlk2 +/+ ) heterozygous ( Tlk1 +/F or Tlk2 +/F ) and homozygous ( Tlk1 F/F or Tlk2 F/F ) mice with primers P1/P2 following deletion of Neo cassette. (E) Left: bright field images of whole mount inguinal mammary glands from 10-week-old Tlk1 F/F ;Tlk2 F/F (control) and Fsp-cre;Tlk1 F/F ;Tlk2 F/F (Tlk1 Δ/Δ ;Tlk2 Δ/Δ ) mice stained with carmine red (bar = 2000 μm). Boxes indicate area of magnification in adjacent panel (bar = 100 μm). Middle: H E stained sections of opposite inguinal gland (bar = 50 μm). Right: representative images of sections of inguinal mammary glands immuno-stained for the proliferation marker Ki67 (brown) and counterstained with hematoxylin (blue) (bar = 50 μm). Boxes indicate area of magnification in adjacent panel (bar = 20 μm). (F) Percent Ki67 positive epithelial cells per 20X field (dots) of tissue sections described in (E). Red bar, median; black bars, interquartile range; pooled counts from 6 control mice or 5 Fsp-cre mice. P value: Mann-Whitney test of medians. (G) Left: Representative images of inguinal mammary gland sections immuno-stained for smooth muscle actin (SMA; brown) and counterstained with hematoxylin (blue) (bar = 50 μm). Insets represent consecutive sections immuno-stained with Ki67 (brown) and counterstained with hematoxylin (blue). Right: Representative images of inguinal mammary gland sections immuno-stained for smooth muscle actin (SMA; red) and Ki67 (brown); counterstained with hematoxylin (blue) (bar = 50 μm). (H) PCR genotyping of mammary fibroblasts derived from Tlk1 F/F and Tlk2 F/F mice with or without Fsp-cre to detect the Flox allele ( Tlk F ; P1/P2 primers), the deletion product formed after Fsp-cre -mediated recombination ( Tlk Δ ; P1/P3 primers) and Fsp-cre. (I) qRT-PCR for Tlk1 or Tlk2 on <t>cDNA</t> derived from fibroblasts isolated from Tlk1 F/F ;Tlk2 F/F mammary glands of control (−) and Fsp-cre positive (+) mice. Expression was normalized to Gapdh. (J) Mammary gland wholemounts from Fsp-cre;Rosa loxP/+ and Rosa loxP/+ (left, inset) mice stained for β-galactosidase. Higher (10×) magnification of a wholemount gland (top-right) and a cross section stained for β-galactosidase and counterstained with nuclear fast red (bottom-right). Str, stromal fibroblasts; lu, lumen; epi, epithelial cells. (K) qRT-PCR for Tlk1 expression (left) on cDNA derived from Tlk1 F/F mammary fibroblasts (MMFs) or mammary epithelial cells (MECs) lacking (−) or containing (+) the Fsp-cre transgene. qRT-PCR for Tlk2 expression (right) on cDNA derived from Tlk2 F/F mammary fibroblasts (MMFs) or mammary epithelial cells (MECs) lacking (−) or containing (+) the Fsp-cre transgene. Expression was normalized to Gapdh. .
    Complementary Dna Cdna Templates, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 88/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 88 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    complementary dna cdna templates - by Bioz Stars, 2020-07
    88/100 stars
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    88
    Quanta Biosciences complementary dna cdna template
    In vivo deletion of Tlk1 and Tlk2 in mouse mammary fibroblasts increases proliferation of adjacent ductal epithelium (A, B) Schematic overview of the targeting vector used to generate the Tlk1 and Tlk2 conditional knock-out mice. Line 1: partial genomic locus of mouse Tlk1 or Tlk2 gene with exons represented as dark gray boxes and Southern probe as light gray bar. Line 2: targeting vector with exons 9 and 10 in Tlk1 and exon 11 in Tlk2 flanked by loxP elements (black triangles), the LacZ reporter cassette and neomycin (Neo) resistance cassette flanked by FRT sites (white triangles). Line 3: targeted genomic locus (Neo). Line 4: Neo cassette was removed by Actin-Flpe mediated recombination ( Flox (F) ). Black arrows P1–P3 indicate location of genotyping primers. Diagram not to scale. (C, D) Southern blot (left panels) of ES cells confirming targeting of Tlk1 or Tlk2 genomic loci respectively. <t>PCR</t> genotyping of genomic DNA (right panels) from WT ( Tlk1 +/+ or Tlk2 +/+ ) heterozygous ( Tlk1 +/F or Tlk2 +/F ) and homozygous ( Tlk1 F/F or Tlk2 F/F ) mice with primers P1/P2 following deletion of Neo cassette. (E) Left: bright field images of whole mount inguinal mammary glands from 10-week-old Tlk1 F/F ;Tlk2 F/F (control) and Fsp-cre;Tlk1 F/F ;Tlk2 F/F (Tlk1 Δ/Δ ;Tlk2 Δ/Δ ) mice stained with carmine red (bar = 2000 μm). Boxes indicate area of magnification in adjacent panel (bar = 100 μm). Middle: H E stained sections of opposite inguinal gland (bar = 50 μm). Right: representative images of sections of inguinal mammary glands immuno-stained for the proliferation marker Ki67 (brown) and counterstained with hematoxylin (blue) (bar = 50 μm). Boxes indicate area of magnification in adjacent panel (bar = 20 μm). (F) Percent Ki67 positive epithelial cells per 20X field (dots) of tissue sections described in (E). Red bar, median; black bars, interquartile range; pooled counts from 6 control mice or 5 Fsp-cre mice. P value: Mann-Whitney test of medians. (G) Left: Representative images of inguinal mammary gland sections immuno-stained for smooth muscle actin (SMA; brown) and counterstained with hematoxylin (blue) (bar = 50 μm). Insets represent consecutive sections immuno-stained with Ki67 (brown) and counterstained with hematoxylin (blue). Right: Representative images of inguinal mammary gland sections immuno-stained for smooth muscle actin (SMA; red) and Ki67 (brown); counterstained with hematoxylin (blue) (bar = 50 μm). (H) PCR genotyping of mammary fibroblasts derived from Tlk1 F/F and Tlk2 F/F mice with or without Fsp-cre to detect the Flox allele ( Tlk F ; P1/P2 primers), the deletion product formed after Fsp-cre -mediated recombination ( Tlk Δ ; P1/P3 primers) and Fsp-cre. (I) qRT-PCR for Tlk1 or Tlk2 on <t>cDNA</t> derived from fibroblasts isolated from Tlk1 F/F ;Tlk2 F/F mammary glands of control (−) and Fsp-cre positive (+) mice. Expression was normalized to Gapdh. (J) Mammary gland wholemounts from Fsp-cre;Rosa loxP/+ and Rosa loxP/+ (left, inset) mice stained for β-galactosidase. Higher (10×) magnification of a wholemount gland (top-right) and a cross section stained for β-galactosidase and counterstained with nuclear fast red (bottom-right). Str, stromal fibroblasts; lu, lumen; epi, epithelial cells. (K) qRT-PCR for Tlk1 expression (left) on cDNA derived from Tlk1 F/F mammary fibroblasts (MMFs) or mammary epithelial cells (MECs) lacking (−) or containing (+) the Fsp-cre transgene. qRT-PCR for Tlk2 expression (right) on cDNA derived from Tlk2 F/F mammary fibroblasts (MMFs) or mammary epithelial cells (MECs) lacking (−) or containing (+) the Fsp-cre transgene. Expression was normalized to Gapdh. .
    Complementary Dna Cdna Template, supplied by Quanta Biosciences, used in various techniques. Bioz Stars score: 88/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    TaKaRa complementary dna cdna templates
    Comparison of the amplification efficiency and time of real-time <t>PCR</t> and RT-LAMP. a Standard curves generated by linear regression analysis of TTP of LAMP and Ct of real-time PCR measured for each amplification versus the log 10 number of DENV-1 RNA or <t>cDNA</t> copies for each standard dilution. b Standard curves generated by linear regression analysis of the time for each amplification versus the log 10 number of DENV-1 RNA or cDNA copies each standard dilution. Data are shown as mean ± standard error of the mean (S.E.M) at least three independent experiments. Ct, cycle threshold; TTP, time-to-positive
    Complementary Dna Cdna Templates, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Roche complementary dna cdna template
    Gene expression levels of several markers related to plant immunity in Nicotiana benthamiana leaves transiently expressing HaSSP30 for 1, 2, and 3 days. Nicotiana benthamiana leaves were infiltrated with Agrobacterium tumefaciens GV3101 carrying either the pICH86988-empty (Control) or pICH86988-HaSSP30 (HaSSP30) vector. The plants were incubated in a plant growth room with 12 h/12 h, night/day photoperiods at 20–24 °C with 60% relative humidity (RH). The leaves were harvested after 1, 2, and 3 days post-infiltration (dpi). Total RNA was extracted and reverse transcribed into <t>cDNA.</t> The <t>qPCR</t> was performed on the cDNA using specific primers for several genes related to activation of plant immunity. Two reference genes were used to normalize the data, which are presented as normalized relative quantities scaled to control. Error bars indicated the standard deviation (SD) of 3 independent replicates (n = 3). Two-tailed Student t-test was used to compare HaSSP30 versus Control in each day and p-values
    Complementary Dna Cdna Template, supplied by Roche, used in various techniques. Bioz Stars score: 88/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa template complementary dna cdna
    The DNA, <t>cDNA</t> nucleotide sequence and deduced amino acid sequence of <t>VpSBP16</t> from V . pesudoreticulata ( A ) and Exon-intron structures of VpSBP16 ( B ). The SBP domain is shown in red and the two zinc-binding sites of the C2HCH type (zinc finger 1 and zinc finger 2) are indicated with green and yellow boxes. The conserved basic amino acids of the nuclear location signal are shaded in dark grey.
    Template Complementary Dna Cdna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Toyobo template complementary dna
    The DNA, <t>cDNA</t> nucleotide sequence and deduced amino acid sequence of <t>VpSBP16</t> from V . pesudoreticulata ( A ) and Exon-intron structures of VpSBP16 ( B ). The SBP domain is shown in red and the two zinc-binding sites of the C2HCH type (zinc finger 1 and zinc finger 2) are indicated with green and yellow boxes. The conserved basic amino acids of the nuclear location signal are shaded in dark grey.
    Template Complementary Dna, 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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    93
    Roche cdna templates
    <t>cDNA</t> and putative amino acid sequences of B. mori Ras. (A) Ras1 (accession number: AB3892674), (B) Ras2 (AB3892674) and (C) Ras3 (AB3892674). Amino acids which are important for GTP/Mg 2+ binding are boxed. The C terminal isoprenylation site is underlined. Glycine residues substituted by valine in <t>BmRas-GFP</t> fusion proteins are indicated by arrowheads.
    Cdna Templates, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 755 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Bio-Rad first strand complementary dna cdna template
    <t>cDNA</t> and putative amino acid sequences of B. mori Ras. (A) Ras1 (accession number: AB3892674), (B) Ras2 (AB3892674) and (C) Ras3 (AB3892674). Amino acids which are important for GTP/Mg 2+ binding are boxed. The C terminal isoprenylation site is underlined. Glycine residues substituted by valine in <t>BmRas-GFP</t> fusion proteins are indicated by arrowheads.
    First Strand Complementary Dna Cdna Template, supplied by Bio-Rad, 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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    90
    Millipore complementary template dna strand oligonucleotide
    <t>cDNA</t> and putative amino acid sequences of B. mori Ras. (A) Ras1 (accession number: AB3892674), (B) Ras2 (AB3892674) and (C) Ras3 (AB3892674). Amino acids which are important for GTP/Mg 2+ binding are boxed. The C terminal isoprenylation site is underlined. Glycine residues substituted by valine in <t>BmRas-GFP</t> fusion proteins are indicated by arrowheads.
    Complementary Template Dna Strand Oligonucleotide, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Bio-Synthesis Inc solution synthetic complementary dna cdna templates
    <t>cDNA</t> and putative amino acid sequences of B. mori Ras. (A) Ras1 (accession number: AB3892674), (B) Ras2 (AB3892674) and (C) Ras3 (AB3892674). Amino acids which are important for GTP/Mg 2+ binding are boxed. The C terminal isoprenylation site is underlined. Glycine residues substituted by valine in <t>BmRas-GFP</t> fusion proteins are indicated by arrowheads.
    Solution Synthetic Complementary Dna Cdna Templates, supplied by Bio-Synthesis Inc, 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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    94
    TaKaRa complementary dna template generated by the reverse transcription kit takara
    <t>cDNA</t> and putative amino acid sequences of B. mori Ras. (A) Ras1 (accession number: AB3892674), (B) Ras2 (AB3892674) and (C) Ras3 (AB3892674). Amino acids which are important for GTP/Mg 2+ binding are boxed. The C terminal isoprenylation site is underlined. Glycine residues substituted by valine in <t>BmRas-GFP</t> fusion proteins are indicated by arrowheads.
    Complementary Dna Template Generated By The Reverse Transcription Kit Takara, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    OriGene cdna template
    <t>cDNA</t> and putative amino acid sequences of B. mori Ras. (A) Ras1 (accession number: AB3892674), (B) Ras2 (AB3892674) and (C) Ras3 (AB3892674). Amino acids which are important for GTP/Mg 2+ binding are boxed. The C terminal isoprenylation site is underlined. Glycine residues substituted by valine in <t>BmRas-GFP</t> fusion proteins are indicated by arrowheads.
    Cdna Template, supplied by OriGene, used in various techniques. Bioz Stars score: 93/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Expression levels of XYLP mRNA in different human tissues. Multiple tissue cDNA panels (Clontech) were used to perform real time PCR analyses. Levels of XYLP ( open bars ) and FAM20B ( closed bars ) mRNAs were normalized to those of glyceraldehyde-3-phosphate

    Journal: The Journal of Biological Chemistry

    Article Title: Identification of Phosphatase That Dephosphorylates Xylose in the Glycosaminoglycan-Protein Linkage Region of Proteoglycans *

    doi: 10.1074/jbc.M113.520536

    Figure Lengend Snippet: Expression levels of XYLP mRNA in different human tissues. Multiple tissue cDNA panels (Clontech) were used to perform real time PCR analyses. Levels of XYLP ( open bars ) and FAM20B ( closed bars ) mRNAs were normalized to those of glyceraldehyde-3-phosphate

    Article Snippet: PCR was used to amplify a cDNA fragment predicted to encode a truncated form of the putative phosphatase (XYLP) lacking the first 37 N-terminal amino acids; the template cDNA was obtained from Open Biosystems (MHS1010-7508558, corresponding to the human acid phosphatase-like-2 (ACPL2) cDNA in GenBankTM ); the 5′-primer (5′-GAAGATCTGGAATGAGTAGCAAGAGTCGA-3′) contained an in-frame BglII site, and the 3′-primer (5′-GAAGATCTGTGGACCTTTCCCTATGCTCT-3′) contained a BglII site located 38 bp downstream of the predicted stop codon.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    In vivo deletion of Tlk1 and Tlk2 in mouse mammary fibroblasts increases proliferation of adjacent ductal epithelium (A, B) Schematic overview of the targeting vector used to generate the Tlk1 and Tlk2 conditional knock-out mice. Line 1: partial genomic locus of mouse Tlk1 or Tlk2 gene with exons represented as dark gray boxes and Southern probe as light gray bar. Line 2: targeting vector with exons 9 and 10 in Tlk1 and exon 11 in Tlk2 flanked by loxP elements (black triangles), the LacZ reporter cassette and neomycin (Neo) resistance cassette flanked by FRT sites (white triangles). Line 3: targeted genomic locus (Neo). Line 4: Neo cassette was removed by Actin-Flpe mediated recombination ( Flox (F) ). Black arrows P1–P3 indicate location of genotyping primers. Diagram not to scale. (C, D) Southern blot (left panels) of ES cells confirming targeting of Tlk1 or Tlk2 genomic loci respectively. PCR genotyping of genomic DNA (right panels) from WT ( Tlk1 +/+ or Tlk2 +/+ ) heterozygous ( Tlk1 +/F or Tlk2 +/F ) and homozygous ( Tlk1 F/F or Tlk2 F/F ) mice with primers P1/P2 following deletion of Neo cassette. (E) Left: bright field images of whole mount inguinal mammary glands from 10-week-old Tlk1 F/F ;Tlk2 F/F (control) and Fsp-cre;Tlk1 F/F ;Tlk2 F/F (Tlk1 Δ/Δ ;Tlk2 Δ/Δ ) mice stained with carmine red (bar = 2000 μm). Boxes indicate area of magnification in adjacent panel (bar = 100 μm). Middle: H E stained sections of opposite inguinal gland (bar = 50 μm). Right: representative images of sections of inguinal mammary glands immuno-stained for the proliferation marker Ki67 (brown) and counterstained with hematoxylin (blue) (bar = 50 μm). Boxes indicate area of magnification in adjacent panel (bar = 20 μm). (F) Percent Ki67 positive epithelial cells per 20X field (dots) of tissue sections described in (E). Red bar, median; black bars, interquartile range; pooled counts from 6 control mice or 5 Fsp-cre mice. P value: Mann-Whitney test of medians. (G) Left: Representative images of inguinal mammary gland sections immuno-stained for smooth muscle actin (SMA; brown) and counterstained with hematoxylin (blue) (bar = 50 μm). Insets represent consecutive sections immuno-stained with Ki67 (brown) and counterstained with hematoxylin (blue). Right: Representative images of inguinal mammary gland sections immuno-stained for smooth muscle actin (SMA; red) and Ki67 (brown); counterstained with hematoxylin (blue) (bar = 50 μm). (H) PCR genotyping of mammary fibroblasts derived from Tlk1 F/F and Tlk2 F/F mice with or without Fsp-cre to detect the Flox allele ( Tlk F ; P1/P2 primers), the deletion product formed after Fsp-cre -mediated recombination ( Tlk Δ ; P1/P3 primers) and Fsp-cre. (I) qRT-PCR for Tlk1 or Tlk2 on cDNA derived from fibroblasts isolated from Tlk1 F/F ;Tlk2 F/F mammary glands of control (−) and Fsp-cre positive (+) mice. Expression was normalized to Gapdh. (J) Mammary gland wholemounts from Fsp-cre;Rosa loxP/+ and Rosa loxP/+ (left, inset) mice stained for β-galactosidase. Higher (10×) magnification of a wholemount gland (top-right) and a cross section stained for β-galactosidase and counterstained with nuclear fast red (bottom-right). Str, stromal fibroblasts; lu, lumen; epi, epithelial cells. (K) qRT-PCR for Tlk1 expression (left) on cDNA derived from Tlk1 F/F mammary fibroblasts (MMFs) or mammary epithelial cells (MECs) lacking (−) or containing (+) the Fsp-cre transgene. qRT-PCR for Tlk2 expression (right) on cDNA derived from Tlk2 F/F mammary fibroblasts (MMFs) or mammary epithelial cells (MECs) lacking (−) or containing (+) the Fsp-cre transgene. Expression was normalized to Gapdh. .

    Journal: Developmental cell

    Article Title: Discovery of stromal regulatory networks that suppress Ras-sensitized epithelial cell proliferation

    doi: 10.1016/j.devcel.2017.04.024

    Figure Lengend Snippet: In vivo deletion of Tlk1 and Tlk2 in mouse mammary fibroblasts increases proliferation of adjacent ductal epithelium (A, B) Schematic overview of the targeting vector used to generate the Tlk1 and Tlk2 conditional knock-out mice. Line 1: partial genomic locus of mouse Tlk1 or Tlk2 gene with exons represented as dark gray boxes and Southern probe as light gray bar. Line 2: targeting vector with exons 9 and 10 in Tlk1 and exon 11 in Tlk2 flanked by loxP elements (black triangles), the LacZ reporter cassette and neomycin (Neo) resistance cassette flanked by FRT sites (white triangles). Line 3: targeted genomic locus (Neo). Line 4: Neo cassette was removed by Actin-Flpe mediated recombination ( Flox (F) ). Black arrows P1–P3 indicate location of genotyping primers. Diagram not to scale. (C, D) Southern blot (left panels) of ES cells confirming targeting of Tlk1 or Tlk2 genomic loci respectively. PCR genotyping of genomic DNA (right panels) from WT ( Tlk1 +/+ or Tlk2 +/+ ) heterozygous ( Tlk1 +/F or Tlk2 +/F ) and homozygous ( Tlk1 F/F or Tlk2 F/F ) mice with primers P1/P2 following deletion of Neo cassette. (E) Left: bright field images of whole mount inguinal mammary glands from 10-week-old Tlk1 F/F ;Tlk2 F/F (control) and Fsp-cre;Tlk1 F/F ;Tlk2 F/F (Tlk1 Δ/Δ ;Tlk2 Δ/Δ ) mice stained with carmine red (bar = 2000 μm). Boxes indicate area of magnification in adjacent panel (bar = 100 μm). Middle: H E stained sections of opposite inguinal gland (bar = 50 μm). Right: representative images of sections of inguinal mammary glands immuno-stained for the proliferation marker Ki67 (brown) and counterstained with hematoxylin (blue) (bar = 50 μm). Boxes indicate area of magnification in adjacent panel (bar = 20 μm). (F) Percent Ki67 positive epithelial cells per 20X field (dots) of tissue sections described in (E). Red bar, median; black bars, interquartile range; pooled counts from 6 control mice or 5 Fsp-cre mice. P value: Mann-Whitney test of medians. (G) Left: Representative images of inguinal mammary gland sections immuno-stained for smooth muscle actin (SMA; brown) and counterstained with hematoxylin (blue) (bar = 50 μm). Insets represent consecutive sections immuno-stained with Ki67 (brown) and counterstained with hematoxylin (blue). Right: Representative images of inguinal mammary gland sections immuno-stained for smooth muscle actin (SMA; red) and Ki67 (brown); counterstained with hematoxylin (blue) (bar = 50 μm). (H) PCR genotyping of mammary fibroblasts derived from Tlk1 F/F and Tlk2 F/F mice with or without Fsp-cre to detect the Flox allele ( Tlk F ; P1/P2 primers), the deletion product formed after Fsp-cre -mediated recombination ( Tlk Δ ; P1/P3 primers) and Fsp-cre. (I) qRT-PCR for Tlk1 or Tlk2 on cDNA derived from fibroblasts isolated from Tlk1 F/F ;Tlk2 F/F mammary glands of control (−) and Fsp-cre positive (+) mice. Expression was normalized to Gapdh. (J) Mammary gland wholemounts from Fsp-cre;Rosa loxP/+ and Rosa loxP/+ (left, inset) mice stained for β-galactosidase. Higher (10×) magnification of a wholemount gland (top-right) and a cross section stained for β-galactosidase and counterstained with nuclear fast red (bottom-right). Str, stromal fibroblasts; lu, lumen; epi, epithelial cells. (K) qRT-PCR for Tlk1 expression (left) on cDNA derived from Tlk1 F/F mammary fibroblasts (MMFs) or mammary epithelial cells (MECs) lacking (−) or containing (+) the Fsp-cre transgene. qRT-PCR for Tlk2 expression (right) on cDNA derived from Tlk2 F/F mammary fibroblasts (MMFs) or mammary epithelial cells (MECs) lacking (−) or containing (+) the Fsp-cre transgene. Expression was normalized to Gapdh. .

    Article Snippet: Each PCR contained 1.5 L of cDNA template and primers at a concentration of 100 nM in a final volume of 20 L of SYBR Green reaction mix (Bio-Rad).

    Techniques: In Vivo, Plasmid Preparation, Knock-Out, Mouse Assay, Southern Blot, Polymerase Chain Reaction, Staining, Marker, MANN-WHITNEY, Derivative Assay, Quantitative RT-PCR, Isolation, Expressing

    Comparison of the amplification efficiency and time of real-time PCR and RT-LAMP. a Standard curves generated by linear regression analysis of TTP of LAMP and Ct of real-time PCR measured for each amplification versus the log 10 number of DENV-1 RNA or cDNA copies for each standard dilution. b Standard curves generated by linear regression analysis of the time for each amplification versus the log 10 number of DENV-1 RNA or cDNA copies each standard dilution. Data are shown as mean ± standard error of the mean (S.E.M) at least three independent experiments. Ct, cycle threshold; TTP, time-to-positive

    Journal: BMC Microbiology

    Article Title: Development of reverse-transcription loop-mediated isothermal amplification assay for rapid detection and differentiation of dengue virus serotypes 1–4

    doi: 10.1186/s12866-015-0595-1

    Figure Lengend Snippet: Comparison of the amplification efficiency and time of real-time PCR and RT-LAMP. a Standard curves generated by linear regression analysis of TTP of LAMP and Ct of real-time PCR measured for each amplification versus the log 10 number of DENV-1 RNA or cDNA copies for each standard dilution. b Standard curves generated by linear regression analysis of the time for each amplification versus the log 10 number of DENV-1 RNA or cDNA copies each standard dilution. Data are shown as mean ± standard error of the mean (S.E.M) at least three independent experiments. Ct, cycle threshold; TTP, time-to-positive

    Article Snippet: Real-time PCR was performed in a 25 μL reaction containing 1 μL cDNA templates, 10 μL 2× SYBR® Premix Ex Taq™(TaKaRa), 2 μL 10 μM F3, 2 μL 10 μM B3, and nuclease-free ddH2O.

    Techniques: Amplification, Real-time Polymerase Chain Reaction, Generated

    Gene expression levels of several markers related to plant immunity in Nicotiana benthamiana leaves transiently expressing HaSSP30 for 1, 2, and 3 days. Nicotiana benthamiana leaves were infiltrated with Agrobacterium tumefaciens GV3101 carrying either the pICH86988-empty (Control) or pICH86988-HaSSP30 (HaSSP30) vector. The plants were incubated in a plant growth room with 12 h/12 h, night/day photoperiods at 20–24 °C with 60% relative humidity (RH). The leaves were harvested after 1, 2, and 3 days post-infiltration (dpi). Total RNA was extracted and reverse transcribed into cDNA. The qPCR was performed on the cDNA using specific primers for several genes related to activation of plant immunity. Two reference genes were used to normalize the data, which are presented as normalized relative quantities scaled to control. Error bars indicated the standard deviation (SD) of 3 independent replicates (n = 3). Two-tailed Student t-test was used to compare HaSSP30 versus Control in each day and p-values

    Journal: Scientific Reports

    Article Title: Small secreted proteins from the necrotrophic conifer pathogen Heterobasidion annosum s.l. (HaSSPs) induce cell death in Nicotiana benthamiana

    doi: 10.1038/s41598-017-08010-0

    Figure Lengend Snippet: Gene expression levels of several markers related to plant immunity in Nicotiana benthamiana leaves transiently expressing HaSSP30 for 1, 2, and 3 days. Nicotiana benthamiana leaves were infiltrated with Agrobacterium tumefaciens GV3101 carrying either the pICH86988-empty (Control) or pICH86988-HaSSP30 (HaSSP30) vector. The plants were incubated in a plant growth room with 12 h/12 h, night/day photoperiods at 20–24 °C with 60% relative humidity (RH). The leaves were harvested after 1, 2, and 3 days post-infiltration (dpi). Total RNA was extracted and reverse transcribed into cDNA. The qPCR was performed on the cDNA using specific primers for several genes related to activation of plant immunity. Two reference genes were used to normalize the data, which are presented as normalized relative quantities scaled to control. Error bars indicated the standard deviation (SD) of 3 independent replicates (n = 3). Two-tailed Student t-test was used to compare HaSSP30 versus Control in each day and p-values

    Article Snippet: The qPCR reaction mixture was assembled with 5.5 µl cDNA template (1:15 dilution), 1 µl forward primer, 1 µl reverse primer, and 7.5 µl 2X LightCycler 480 SYBR Green I Master (Roche, Finland).

    Techniques: Expressing, Plasmid Preparation, Incubation, Real-time Polymerase Chain Reaction, Activation Assay, Standard Deviation, Two Tailed Test

    KGFR and KGF in pancreatic cancer cell lines. A: RT-PCR analysis of KGFR and KGF mRNA in pancreatic cancer cell lines. Total RNA was extracted from PANC-1; MIA PaCa-2; KLM-1; Capan-1; and PK-1, -8, -9, and -59 cells and used to perform cDNA synthesis and PCR analysis. KGFR mRNA (150 bp) was detected in all cell lines (top). KGF mRNA (163-bp) was detected in all pancreatic cancer cell lines, except for MIA PaCa-2 cells (middle). β-Actin mRNA (163 bp) served as a loading control (bottom). B and C: Q-PCR analysis of KGFR and KGF mRNA in pancreatic cancer cell lines. KGFR mRNA was expressed at variable levels in all pancreatic cancer cell lines. KGF mRNA was not detected in MIA PaCa-2 cells and was expressed at variable levels in the other pancreatic cancer cell lines. Results are expressed as KGFR/β-actin and KGF/β-actin. Gene expression measurements were performed in triplicate. Bars represent the mean ± SE. D: Western blot analysis of KGFR in pancreatic cancer cell lines. A band corresponding to the 105-kd KGFR protein was detected in all pancreatic cancer cell lines (top). β-Actin served as a loading control (bottom). E: Immunofluorescent analysis of KGFR expression in MIA PaCa-2 cells. KGFR immunoreactivity was detected in the cytoplasm and/or on the membrane of cancer cells. In some cases, a strong KGFR signal was detected at the cell membrane ( arrowheads ). Bar = 50 μm.

    Journal: The American Journal of Pathology

    Article Title: Enhanced Expression of Keratinocyte Growth Factor and Its Receptor Correlates with Venous Invasion in Pancreatic Cancer

    doi: 10.2353/ajpath.2007.060935

    Figure Lengend Snippet: KGFR and KGF in pancreatic cancer cell lines. A: RT-PCR analysis of KGFR and KGF mRNA in pancreatic cancer cell lines. Total RNA was extracted from PANC-1; MIA PaCa-2; KLM-1; Capan-1; and PK-1, -8, -9, and -59 cells and used to perform cDNA synthesis and PCR analysis. KGFR mRNA (150 bp) was detected in all cell lines (top). KGF mRNA (163-bp) was detected in all pancreatic cancer cell lines, except for MIA PaCa-2 cells (middle). β-Actin mRNA (163 bp) served as a loading control (bottom). B and C: Q-PCR analysis of KGFR and KGF mRNA in pancreatic cancer cell lines. KGFR mRNA was expressed at variable levels in all pancreatic cancer cell lines. KGF mRNA was not detected in MIA PaCa-2 cells and was expressed at variable levels in the other pancreatic cancer cell lines. Results are expressed as KGFR/β-actin and KGF/β-actin. Gene expression measurements were performed in triplicate. Bars represent the mean ± SE. D: Western blot analysis of KGFR in pancreatic cancer cell lines. A band corresponding to the 105-kd KGFR protein was detected in all pancreatic cancer cell lines (top). β-Actin served as a loading control (bottom). E: Immunofluorescent analysis of KGFR expression in MIA PaCa-2 cells. KGFR immunoreactivity was detected in the cytoplasm and/or on the membrane of cancer cells. In some cases, a strong KGFR signal was detected at the cell membrane ( arrowheads ). Bar = 50 μm.

    Article Snippet: PCR reaction mixture containing 2 μl of template cDNA, 3 mmol/L MgCl2 , and 0.5 μmol/L of primers, and LightCycler-FastStart DNA Master SYBR Green I mix was applied into a capillary tube (Roche).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Expressing, Western Blot

    The DNA, cDNA nucleotide sequence and deduced amino acid sequence of VpSBP16 from V . pesudoreticulata ( A ) and Exon-intron structures of VpSBP16 ( B ). The SBP domain is shown in red and the two zinc-binding sites of the C2HCH type (zinc finger 1 and zinc finger 2) are indicated with green and yellow boxes. The conserved basic amino acids of the nuclear location signal are shaded in dark grey.

    Journal: International Journal of Molecular Sciences

    Article Title: Overexpression of a SBP-Box Gene (VpSBP16) from Chinese Wild Vitis Species in Arabidopsis Improves Salinity and Drought Stress Tolerance

    doi: 10.3390/ijms19040940

    Figure Lengend Snippet: The DNA, cDNA nucleotide sequence and deduced amino acid sequence of VpSBP16 from V . pesudoreticulata ( A ) and Exon-intron structures of VpSBP16 ( B ). The SBP domain is shown in red and the two zinc-binding sites of the C2HCH type (zinc finger 1 and zinc finger 2) are indicated with green and yellow boxes. The conserved basic amino acids of the nuclear location signal are shaded in dark grey.

    Article Snippet: A pair of gene-specific primers (VpSBP16 -F1 and VpSBP16 -R1) ( ) were used to amplify the predicted VpSBP16 ORF from the cDNA template with Taq DNA polymerase (TaKaRa Biotechnology, Dalian, China) and the following cycling program: 94 °C for 3 min, 35 cycles at 94 °C for 30 s, 58 °C for 30 s and 72 °C for 2 min; and extension at 72 °C for 10 min.

    Techniques: Sequencing, Binding Assay

    Cloning of VpSBP16 from V . pesudoreticulata . ( A ) PCR amplification of the full-length VpSBP16 cDNA and ( B ) DNA from Vitis pseudoreticulata . M1: DNA marker DL5000; M2: DNA marker λ-Hind III; 1: VpSBP16 PCR product from cDNA; 2 and 3: VpSBP16 PCR product.

    Journal: International Journal of Molecular Sciences

    Article Title: Overexpression of a SBP-Box Gene (VpSBP16) from Chinese Wild Vitis Species in Arabidopsis Improves Salinity and Drought Stress Tolerance

    doi: 10.3390/ijms19040940

    Figure Lengend Snippet: Cloning of VpSBP16 from V . pesudoreticulata . ( A ) PCR amplification of the full-length VpSBP16 cDNA and ( B ) DNA from Vitis pseudoreticulata . M1: DNA marker DL5000; M2: DNA marker λ-Hind III; 1: VpSBP16 PCR product from cDNA; 2 and 3: VpSBP16 PCR product.

    Article Snippet: A pair of gene-specific primers (VpSBP16 -F1 and VpSBP16 -R1) ( ) were used to amplify the predicted VpSBP16 ORF from the cDNA template with Taq DNA polymerase (TaKaRa Biotechnology, Dalian, China) and the following cycling program: 94 °C for 3 min, 35 cycles at 94 °C for 30 s, 58 °C for 30 s and 72 °C for 2 min; and extension at 72 °C for 10 min.

    Techniques: Clone Assay, Polymerase Chain Reaction, Amplification, Marker

    Exon/intron structure and alternative mRNA transcripts of mouse GAD1 gene. The new arrangement of mouse GAD1 exons and introns as determined after the analysis of genomic and cDNA sequence data using bioinformatics, 3′-RACE, RT-PCR, cloning and sequencing. Exons are shown as numbered boxes (red numbers represent alternatively spliced exons) and introns as lines. Large boxes indicate the coding DNA sequence and the small boxes the 5′- and 3′-untranslated regions. The red arrowheads are showing the locations of the alternative promoters. The schematic representation of GAD1 splicing isoforms in relation to the gene is shown below the gene structure. The length in base pairs and the position of start and stop codons are indicated above each isoform. Diagrams are not to scale.

    Journal: BMC Neuroscience

    Article Title: Glutamic acid decarboxylase 1 alternative splicing isoforms: characterization, expression and quantification in the mouse brain

    doi: 10.1186/1471-2202-15-114

    Figure Lengend Snippet: Exon/intron structure and alternative mRNA transcripts of mouse GAD1 gene. The new arrangement of mouse GAD1 exons and introns as determined after the analysis of genomic and cDNA sequence data using bioinformatics, 3′-RACE, RT-PCR, cloning and sequencing. Exons are shown as numbered boxes (red numbers represent alternatively spliced exons) and introns as lines. Large boxes indicate the coding DNA sequence and the small boxes the 5′- and 3′-untranslated regions. The red arrowheads are showing the locations of the alternative promoters. The schematic representation of GAD1 splicing isoforms in relation to the gene is shown below the gene structure. The length in base pairs and the position of start and stop codons are indicated above each isoform. Diagrams are not to scale.

    Article Snippet: The PCR mix (25 μl) contained 2.5 μl template cDNA, 1 μM forward and reverse primers, 200 μM dNTPs (Takara), and 0.5 U Ex Taq HS (Takara).

    Techniques: Sequencing, Reverse Transcription Polymerase Chain Reaction, Clone Assay

    Expression analysis of mouse GAD1 splicing isoforms in adult brain. (A) RT-PCR analysis of the expression of GAD1 mRNA splicing isoforms in adult mouse brain by using a forward primer either in Exon1 or Exon2 and a specific reverse primer for each transcript. (B) Analysis of the expression of Isoforms 1 to 6 by using the PCR product of lanes Exon1 and Exon2 in (A) as template for nested PCR with specific primers for each isoform. (C) Gel electrophoresis of Isoforms 3/4 and 5/6 , amplified from mouse brain cDNA using specific forward primer and reverse primer, close to the position of the polyadenylation signal. Plasmid containing Isoforms 1/2 was used as a negative control with the primers for Isoforms 3/4 and 5/6 and as a positive control with primers amplifying the 3′-end of Isoforms 1/2 . (D) Southern blotting of the gel in (C) . The membrane was probed with Probe-GAD1 against 3′-region of Isoforms 1 and 2 . Ma, marker λ/HindIII-ϕX174/HaeIII.

    Journal: BMC Neuroscience

    Article Title: Glutamic acid decarboxylase 1 alternative splicing isoforms: characterization, expression and quantification in the mouse brain

    doi: 10.1186/1471-2202-15-114

    Figure Lengend Snippet: Expression analysis of mouse GAD1 splicing isoforms in adult brain. (A) RT-PCR analysis of the expression of GAD1 mRNA splicing isoforms in adult mouse brain by using a forward primer either in Exon1 or Exon2 and a specific reverse primer for each transcript. (B) Analysis of the expression of Isoforms 1 to 6 by using the PCR product of lanes Exon1 and Exon2 in (A) as template for nested PCR with specific primers for each isoform. (C) Gel electrophoresis of Isoforms 3/4 and 5/6 , amplified from mouse brain cDNA using specific forward primer and reverse primer, close to the position of the polyadenylation signal. Plasmid containing Isoforms 1/2 was used as a negative control with the primers for Isoforms 3/4 and 5/6 and as a positive control with primers amplifying the 3′-end of Isoforms 1/2 . (D) Southern blotting of the gel in (C) . The membrane was probed with Probe-GAD1 against 3′-region of Isoforms 1 and 2 . Ma, marker λ/HindIII-ϕX174/HaeIII.

    Article Snippet: The PCR mix (25 μl) contained 2.5 μl template cDNA, 1 μM forward and reverse primers, 200 μM dNTPs (Takara), and 0.5 U Ex Taq HS (Takara).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Nested PCR, Nucleic Acid Electrophoresis, Amplification, Plasmid Preparation, Negative Control, Positive Control, Southern Blot, Marker

    Expression of GAD1 mRNA splicing isoforms and GAD2 in different mouse tissues and during development. The expression level of Isoforms 1/2 (A) , GAD2 (B) , Isoforms 3/4 (C) , Isoforms 5/6 (D) , Isoforms 7/8 (E) , Isoforms 9/10 (F) are compared by PCR amplification using mouse Multiple Tissue cDNA Panel I and cDNA from pancreas, small intestine, and large intestine as cDNA template. (G) Control PCR reaction to verify the specificity of the primers for Isoforms 1/2, 3/4 and 5/6 . In the control reaction each primer pair was tested with a plasmid containing each full length insert as a template. The template in lanes 1, 4 and 7 was plasmid containing Isoforms 1/2 as template; lanes 2, 5 and 8, plasmid containing Isoforms 5/6 ; and lanes 3, 6, 9 plasmid containing Isoforms 3/4 . Lanes (Ht) heart; (Br) brain; (Sp) spleen; (L) lung; (Li) liver; (Ms) muscle; (K) kidney; (Ts) testis; (E7) 7-day embryo; (E11) 11-day embryo; (E15) 15-day embryo; (E17) 17-day embryo; (P) pancreas; (SI) small intestine; (LI) large intestine; (N) no template control; (−) plasmid containing Isoforms 1/2 is used as template for the amplification; and Ma, marker ϕX174/HaeIII.

    Journal: BMC Neuroscience

    Article Title: Glutamic acid decarboxylase 1 alternative splicing isoforms: characterization, expression and quantification in the mouse brain

    doi: 10.1186/1471-2202-15-114

    Figure Lengend Snippet: Expression of GAD1 mRNA splicing isoforms and GAD2 in different mouse tissues and during development. The expression level of Isoforms 1/2 (A) , GAD2 (B) , Isoforms 3/4 (C) , Isoforms 5/6 (D) , Isoforms 7/8 (E) , Isoforms 9/10 (F) are compared by PCR amplification using mouse Multiple Tissue cDNA Panel I and cDNA from pancreas, small intestine, and large intestine as cDNA template. (G) Control PCR reaction to verify the specificity of the primers for Isoforms 1/2, 3/4 and 5/6 . In the control reaction each primer pair was tested with a plasmid containing each full length insert as a template. The template in lanes 1, 4 and 7 was plasmid containing Isoforms 1/2 as template; lanes 2, 5 and 8, plasmid containing Isoforms 5/6 ; and lanes 3, 6, 9 plasmid containing Isoforms 3/4 . Lanes (Ht) heart; (Br) brain; (Sp) spleen; (L) lung; (Li) liver; (Ms) muscle; (K) kidney; (Ts) testis; (E7) 7-day embryo; (E11) 11-day embryo; (E15) 15-day embryo; (E17) 17-day embryo; (P) pancreas; (SI) small intestine; (LI) large intestine; (N) no template control; (−) plasmid containing Isoforms 1/2 is used as template for the amplification; and Ma, marker ϕX174/HaeIII.

    Article Snippet: The PCR mix (25 μl) contained 2.5 μl template cDNA, 1 μM forward and reverse primers, 200 μM dNTPs (Takara), and 0.5 U Ex Taq HS (Takara).

    Techniques: Expressing, Polymerase Chain Reaction, Amplification, Plasmid Preparation, Mass Spectrometry, Marker

    cDNA and putative amino acid sequences of B. mori Ras. (A) Ras1 (accession number: AB3892674), (B) Ras2 (AB3892674) and (C) Ras3 (AB3892674). Amino acids which are important for GTP/Mg 2+ binding are boxed. The C terminal isoprenylation site is underlined. Glycine residues substituted by valine in BmRas-GFP fusion proteins are indicated by arrowheads.

    Journal: PLoS ONE

    Article Title: Identification and Expression Analysis of Ras Gene in Silkworm, Bombyx mori

    doi: 10.1371/journal.pone.0008030

    Figure Lengend Snippet: cDNA and putative amino acid sequences of B. mori Ras. (A) Ras1 (accession number: AB3892674), (B) Ras2 (AB3892674) and (C) Ras3 (AB3892674). Amino acids which are important for GTP/Mg 2+ binding are boxed. The C terminal isoprenylation site is underlined. Glycine residues substituted by valine in BmRas-GFP fusion proteins are indicated by arrowheads.

    Article Snippet: The transcripts of Bmras , the same as Bmrp49 , in prepared cDNA templates were quantified on a real-time thermal cycler, LightCycler 480 Real-Time PCR System (Roche Diagnostics, Indianapolis, IN, USA). qRT-PCR was carried out in 20 µl reaction volume containing 2 µl template cDNA (equivalent to 5 ng total RNA) or standard cDNA, 1 x SYBR Premix EX Taq (TaKaRa) and each primer.

    Techniques: Binding Assay