cdna template Takara Search Results


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  • 94
    TaKaRa complementary dna template generated by the reverse transcription kit takara
    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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    TaKaRa complementary dna cdna
    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, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 8915 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa complementary dna cdna synthesis
    Effect of Cymbidium ethanol extract (CYM) on 2,4-dinitrochlorobenzene- (DNCB-) induced expression of interleukin- (IL-) 4, IL-13, and tumor necrosis factor- (TNF-) α mRNA in atopic dermatitis- (AD-) like mouse model. After inducing AD, 10 mg/mL CYM solution (in 3 : 1 mixture of acetone and olive oil) was applied to the dorsal skin of mice for a total of 6 times over a 2-week period. After euthanasia, dorsal skin lesions were enucleated from mice of all three groups (non-, DNCB, DNCB, and CYM treatment). Total <t>RNA</t> was isolated from extracted tissue of dorsal lesions using Tri-reagent. Total RNA was used as a template for <t>cDNA</t> synthesis, which was performed using a cDNA Synthesis kit. Real-time polymerase chain reaction (qPCR) analysis was carried out using SYBR Green I and a Lightcycler 96 instrument. The qPCR analysis was performed to detect IL-4, IL-13, and TNF- α mRNA expression. Data are mean ± standard deviation (SD, n = 6). ∗ P
    Complementary Dna Cdna Synthesis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 702 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    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 438 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    TaKaRa complementary dna cdna reverse transcription
    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.
    Complementary Dna Cdna Reverse Transcription, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    TaKaRa marathon cdna amplification kit
    Multiple-tissue Northern blots and RT-PCRs of fish connexins. A , Northern blot of carpCx43 <t>mRNA.</t> CarpCx43 mRNA is highest in brain and retina, whereas liver reveals no detectable levels. B , Ethidium bromide-stained agarose gel of A that shows comparative loading of mRNA (18S and 28S band). C , Multiple-tissue RT-PCR of the three zebrafish connexins (ethidium bromide-stained agarose gels). zfCx44.1 is less abundant in brain and retina, with higher levels in lens and heart. zfCx27.5 shows tissue-restricted expression in brain and retina, whereas zfCx55.5 is exclusively expressed in the retina. All <t>cDNA</t> preparations were controlled by PCR using primers specific for β-actin. D , PCR control (with β-actin primers) omitting reverse transcription. No amplification is evident with the exception of the genomic DNA, indicative of lack of genomic DNA contamination in the mRNA samples.
    Marathon Cdna Amplification Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 98/100, based on 2527 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    TaKaRa snx2 cdna
    Construction and characterization of SNX1, SNX1A, <t>SNX2,</t> SNX3, and SNX4 cDNAs. Five human sorting nexins (SNX1, SNX1A, SNX2, SNX3, and SNX4) are depicted. For each full-length <t>cDNA,</t> the nucleotide numbering is shown. Listed below each molecule are the fragments (ESTs, PCR, or 5′-RACE products) used to determine the nucleotide sequences of the cDNAs and to construct full-length cDNAs as detailed in Materials and Methods. One or both strands of each cDNA fragment were sequenced as indicated by the directions of the arrowheads. In every case, both strands of DNA were sequenced from at least one clone.
    Snx2 Cdna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 80/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa cdna synthesis kit
    Construction and characterization of SNX1, SNX1A, <t>SNX2,</t> SNX3, and SNX4 cDNAs. Five human sorting nexins (SNX1, SNX1A, SNX2, SNX3, and SNX4) are depicted. For each full-length <t>cDNA,</t> the nucleotide numbering is shown. Listed below each molecule are the fragments (ESTs, PCR, or 5′-RACE products) used to determine the nucleotide sequences of the cDNAs and to construct full-length cDNAs as detailed in Materials and Methods. One or both strands of each cDNA fragment were sequenced as indicated by the directions of the arrowheads. In every case, both strands of DNA were sequenced from at least one clone.
    Cdna Synthesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 5208 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    TaKaRa mouse cdna template
    Construction and characterization of SNX1, SNX1A, <t>SNX2,</t> SNX3, and SNX4 cDNAs. Five human sorting nexins (SNX1, SNX1A, SNX2, SNX3, and SNX4) are depicted. For each full-length <t>cDNA,</t> the nucleotide numbering is shown. Listed below each molecule are the fragments (ESTs, PCR, or 5′-RACE products) used to determine the nucleotide sequences of the cDNAs and to construct full-length cDNAs as detailed in Materials and Methods. One or both strands of each cDNA fragment were sequenced as indicated by the directions of the arrowheads. In every case, both strands of DNA were sequenced from at least one clone.
    Mouse Cdna Template, supplied by TaKaRa, used in various techniques. Bioz Stars score: 80/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    BioChain Institute cdna templates
    Construction and characterization of SNX1, SNX1A, <t>SNX2,</t> SNX3, and SNX4 cDNAs. Five human sorting nexins (SNX1, SNX1A, SNX2, SNX3, and SNX4) are depicted. For each full-length <t>cDNA,</t> the nucleotide numbering is shown. Listed below each molecule are the fragments (ESTs, PCR, or 5′-RACE products) used to determine the nucleotide sequences of the cDNAs and to construct full-length cDNAs as detailed in Materials and Methods. One or both strands of each cDNA fragment were sequenced as indicated by the directions of the arrowheads. In every case, both strands of DNA were sequenced from at least one clone.
    Cdna Templates, supplied by BioChain Institute, 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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    94
    Roche cdna template
    Standard curves for <t>RT-qPCR</t> assays of G6PDH and EF1 mRNA expression. Standard curve for G6PDH (a) was generated using V . mali <t>cDNA</t> serially (10 9 to 10 2 fg/μL) diluted in host cDNA (10 9 fg/μL). Standard curve for EF1 (b) was generated by Malus × domestica cv. Fuji cDNA diluted serially (10 9 to 10 2 fg/μL) in water. A linear regression curve was calculated for use to quantify RNA levels from plant samples.
    Cdna Template, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 2251 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa smarttm pcr cdna synthesis kit
    Standard curves for <t>RT-qPCR</t> assays of G6PDH and EF1 mRNA expression. Standard curve for G6PDH (a) was generated using V . mali <t>cDNA</t> serially (10 9 to 10 2 fg/μL) diluted in host cDNA (10 9 fg/μL). Standard curve for EF1 (b) was generated by Malus × domestica cv. Fuji cDNA diluted serially (10 9 to 10 2 fg/μL) in water. A linear regression curve was calculated for use to quantify RNA levels from plant samples.
    Smarttm Pcr Cdna Synthesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 135 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    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

    Cloning, amino acid sequence alignment, and an unrooted phylogenetic tree of DFR obtained from deduced amino acid sequences of 13 DFR homologs. ( a ) a DFR cDNA fragment was amplified from V. bellula leaf tissue by RT-PCR. M: DNA marker, 1: DFR cDNA fragment, CT: negative control; ( b ) an unrooted phylogenetic tree was built from amino acid sequences of 13 DFR homologs; ( c ) amino acid sequence alignment were develop from 13 DFR homologs; “*”: the same amino acid in all sequences; “:”: conserved amino acid residues; “.”: half conserved amino acid residues; “######”: potential NADPH/NADH binding domain; amino acids underlined form a potential substrate specificity domain of DFR. AtDFR: Arabidopsis thaliana DFR; CmDFR: Crataegus monogyna DFR; GmDFR1: Glycine max DFR1; GmDFR2: Glycine max DFR2; LjDFR: Lotus japonicas DFR; MdDFR: Malus domestica DFR; MtDFR: Medicago truncatula DFR; NgDFR: Nekemias grossedentata DFR; PaDFR: Prunus avium DFR; PpDFR: Pyrus pyrifolia DFR; PlDFR: Paeonia lactiflora DFR; VvDFR: Vitis vinifera DFR.

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: Molecular Cloning and Functional Characterization of a Dihydroflavonol 4-Reductase from Vitis bellula

    doi: 10.3390/molecules23040861

    Figure Lengend Snippet: Cloning, amino acid sequence alignment, and an unrooted phylogenetic tree of DFR obtained from deduced amino acid sequences of 13 DFR homologs. ( a ) a DFR cDNA fragment was amplified from V. bellula leaf tissue by RT-PCR. M: DNA marker, 1: DFR cDNA fragment, CT: negative control; ( b ) an unrooted phylogenetic tree was built from amino acid sequences of 13 DFR homologs; ( c ) amino acid sequence alignment were develop from 13 DFR homologs; “*”: the same amino acid in all sequences; “:”: conserved amino acid residues; “.”: half conserved amino acid residues; “######”: potential NADPH/NADH binding domain; amino acids underlined form a potential substrate specificity domain of DFR. AtDFR: Arabidopsis thaliana DFR; CmDFR: Crataegus monogyna DFR; GmDFR1: Glycine max DFR1; GmDFR2: Glycine max DFR2; LjDFR: Lotus japonicas DFR; MdDFR: Malus domestica DFR; MtDFR: Medicago truncatula DFR; NgDFR: Nekemias grossedentata DFR; PaDFR: Prunus avium DFR; PpDFR: Pyrus pyrifolia DFR; PlDFR: Paeonia lactiflora DFR; VvDFR: Vitis vinifera DFR.

    Article Snippet: The resulting DNA-free RNA sample was used as a template to synthesize the first strand cDNA with MMLV Reverse transcriptase (Takara, Japan) and oligo (dT)12 primer.

    Techniques: Clone Assay, Sequencing, Amplification, Reverse Transcription Polymerase Chain Reaction, Marker, Negative Control, Binding Assay

    Effect of Cymbidium ethanol extract (CYM) on 2,4-dinitrochlorobenzene- (DNCB-) induced expression of interleukin- (IL-) 4, IL-13, and tumor necrosis factor- (TNF-) α mRNA in atopic dermatitis- (AD-) like mouse model. After inducing AD, 10 mg/mL CYM solution (in 3 : 1 mixture of acetone and olive oil) was applied to the dorsal skin of mice for a total of 6 times over a 2-week period. After euthanasia, dorsal skin lesions were enucleated from mice of all three groups (non-, DNCB, DNCB, and CYM treatment). Total RNA was isolated from extracted tissue of dorsal lesions using Tri-reagent. Total RNA was used as a template for cDNA synthesis, which was performed using a cDNA Synthesis kit. Real-time polymerase chain reaction (qPCR) analysis was carried out using SYBR Green I and a Lightcycler 96 instrument. The qPCR analysis was performed to detect IL-4, IL-13, and TNF- α mRNA expression. Data are mean ± standard deviation (SD, n = 6). ∗ P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Effects of Cymbidium Root Ethanol Extract on Atopic Dermatitis

    doi: 10.1155/2016/5362475

    Figure Lengend Snippet: Effect of Cymbidium ethanol extract (CYM) on 2,4-dinitrochlorobenzene- (DNCB-) induced expression of interleukin- (IL-) 4, IL-13, and tumor necrosis factor- (TNF-) α mRNA in atopic dermatitis- (AD-) like mouse model. After inducing AD, 10 mg/mL CYM solution (in 3 : 1 mixture of acetone and olive oil) was applied to the dorsal skin of mice for a total of 6 times over a 2-week period. After euthanasia, dorsal skin lesions were enucleated from mice of all three groups (non-, DNCB, DNCB, and CYM treatment). Total RNA was isolated from extracted tissue of dorsal lesions using Tri-reagent. Total RNA was used as a template for cDNA synthesis, which was performed using a cDNA Synthesis kit. Real-time polymerase chain reaction (qPCR) analysis was carried out using SYBR Green I and a Lightcycler 96 instrument. The qPCR analysis was performed to detect IL-4, IL-13, and TNF- α mRNA expression. Data are mean ± standard deviation (SD, n = 6). ∗ P

    Article Snippet: Total RNA was used as a template for cDNA synthesis, which was performed using a cDNA Synthesis kit (Takara Bio, Shiga, Japan).

    Techniques: Expressing, Mouse Assay, Isolation, Real-time Polymerase Chain Reaction, SYBR Green Assay, Standard Deviation

    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

    Multiple-tissue Northern blots and RT-PCRs of fish connexins. A , Northern blot of carpCx43 mRNA. CarpCx43 mRNA is highest in brain and retina, whereas liver reveals no detectable levels. B , Ethidium bromide-stained agarose gel of A that shows comparative loading of mRNA (18S and 28S band). C , Multiple-tissue RT-PCR of the three zebrafish connexins (ethidium bromide-stained agarose gels). zfCx44.1 is less abundant in brain and retina, with higher levels in lens and heart. zfCx27.5 shows tissue-restricted expression in brain and retina, whereas zfCx55.5 is exclusively expressed in the retina. All cDNA preparations were controlled by PCR using primers specific for β-actin. D , PCR control (with β-actin primers) omitting reverse transcription. No amplification is evident with the exception of the genomic DNA, indicative of lack of genomic DNA contamination in the mRNA samples.

    Journal: The Journal of Neuroscience

    Article Title: Molecular and Functional Diversity of Neural Connexins in the Retina

    doi: 10.1523/JNEUROSCI.20-22-08331.2000

    Figure Lengend Snippet: Multiple-tissue Northern blots and RT-PCRs of fish connexins. A , Northern blot of carpCx43 mRNA. CarpCx43 mRNA is highest in brain and retina, whereas liver reveals no detectable levels. B , Ethidium bromide-stained agarose gel of A that shows comparative loading of mRNA (18S and 28S band). C , Multiple-tissue RT-PCR of the three zebrafish connexins (ethidium bromide-stained agarose gels). zfCx44.1 is less abundant in brain and retina, with higher levels in lens and heart. zfCx27.5 shows tissue-restricted expression in brain and retina, whereas zfCx55.5 is exclusively expressed in the retina. All cDNA preparations were controlled by PCR using primers specific for β-actin. D , PCR control (with β-actin primers) omitting reverse transcription. No amplification is evident with the exception of the genomic DNA, indicative of lack of genomic DNA contamination in the mRNA samples.

    Article Snippet: In addition to the screening of the genomic zebrafish library, we performed RACE extension using carp mRNA as a template for the Marathon cDNA Amplification Kit (Clontech Laboratories, Heidelberg, Germany).

    Techniques: Northern Blot, Fluorescence In Situ Hybridization, Staining, Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Expressing, Polymerase Chain Reaction, Amplification

    Construction and characterization of SNX1, SNX1A, SNX2, SNX3, and SNX4 cDNAs. Five human sorting nexins (SNX1, SNX1A, SNX2, SNX3, and SNX4) are depicted. For each full-length cDNA, the nucleotide numbering is shown. Listed below each molecule are the fragments (ESTs, PCR, or 5′-RACE products) used to determine the nucleotide sequences of the cDNAs and to construct full-length cDNAs as detailed in Materials and Methods. One or both strands of each cDNA fragment were sequenced as indicated by the directions of the arrowheads. In every case, both strands of DNA were sequenced from at least one clone.

    Journal: Molecular and Cellular Biology

    Article Title: Identification of a Family of Sorting Nexin Molecules and Characterization of Their Association with Receptors

    doi:

    Figure Lengend Snippet: Construction and characterization of SNX1, SNX1A, SNX2, SNX3, and SNX4 cDNAs. Five human sorting nexins (SNX1, SNX1A, SNX2, SNX3, and SNX4) are depicted. For each full-length cDNA, the nucleotide numbering is shown. Listed below each molecule are the fragments (ESTs, PCR, or 5′-RACE products) used to determine the nucleotide sequences of the cDNAs and to construct full-length cDNAs as detailed in Materials and Methods. One or both strands of each cDNA fragment were sequenced as indicated by the directions of the arrowheads. In every case, both strands of DNA were sequenced from at least one clone.

    Article Snippet: A 5′-rapid amplification of cDNA ends (RACE) strategy (Life Technologies, Inc., Gaithersburg, Md.) with total human skeletal muscle as a template was used to complete the 5′ end of the SNX2 cDNA (Clontech).

    Techniques: Polymerase Chain Reaction, Construct

    Standard curves for RT-qPCR assays of G6PDH and EF1 mRNA expression. Standard curve for G6PDH (a) was generated using V . mali cDNA serially (10 9 to 10 2 fg/μL) diluted in host cDNA (10 9 fg/μL). Standard curve for EF1 (b) was generated by Malus × domestica cv. Fuji cDNA diluted serially (10 9 to 10 2 fg/μL) in water. A linear regression curve was calculated for use to quantify RNA levels from plant samples.

    Journal: PLoS ONE

    Article Title: Study on Interactions between the Major Apple Valsa Canker Pathogen Valsa mali and Its Biocontrol Agent Saccharothrix yanglingensis Hhs.015 Using RT-qPCR

    doi: 10.1371/journal.pone.0162174

    Figure Lengend Snippet: Standard curves for RT-qPCR assays of G6PDH and EF1 mRNA expression. Standard curve for G6PDH (a) was generated using V . mali cDNA serially (10 9 to 10 2 fg/μL) diluted in host cDNA (10 9 fg/μL). Standard curve for EF1 (b) was generated by Malus × domestica cv. Fuji cDNA diluted serially (10 9 to 10 2 fg/μL) in water. A linear regression curve was calculated for use to quantify RNA levels from plant samples.

    Article Snippet: RT-qPCR assay Experimental samples were analyzed using a Bio-Rad iQ5, qPCR assay in 25 μL reactions containing 1 μL of cDNA template, 0.4 μL of each primers (0.2 μM), 1 × reaction buffer, 2 2 μM MgCl2 , 0.2 2 μM dNTPs (Roche, Mannheim, Germany), 0.4 U Taq DNA polymerase (Thermo Scientific, Lithuania), 2 μL of cDNA template, and 0.5 mL of 2 × SYBR (Takara, Tokyo, Japan).

    Techniques: Quantitative RT-PCR, Expressing, Generated