cdna template Takara Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher trizol reagent
    Quantification of the mRNA transcriptional levels of EGFP (A) and IL2 (B). Tumor cells in 6-well plate were infected with the indicated rNDV-EGFPs or rNDV-IL2s at MOI of 10. Total <t>RNA</t> in infected cells was extracted using <t>Trizol</t> reagent at 48 hpi, and qRT-PCR was performed to quantify mRNA transcriptional levels of EGFP and IL2, respectively. Transcriptional levels of EGFP and IL2 genes are calculated relative to the β-actin, and the data are expressed as fold changes with SEM (* P
    Trizol Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 604295 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trizol reagent/product/Thermo Fisher
    Average 99 stars, based on 604295 article reviews
    Price from $9.99 to $1999.99
    trizol reagent - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa primescript rt master mix
    Quantification of the mRNA transcriptional levels of EGFP (A) and IL2 (B). Tumor cells in 6-well plate were infected with the indicated rNDV-EGFPs or rNDV-IL2s at MOI of 10. Total <t>RNA</t> in infected cells was extracted using <t>Trizol</t> reagent at 48 hpi, and qRT-PCR was performed to quantify mRNA transcriptional levels of EGFP and IL2, respectively. Transcriptional levels of EGFP and IL2 genes are calculated relative to the β-actin, and the data are expressed as fold changes with SEM (* P
    Primescript Rt Master Mix, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 12772 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primescript rt master mix/product/TaKaRa
    Average 99 stars, based on 12772 article reviews
    Price from $9.99 to $1999.99
    primescript rt master mix - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa pcr amplification
    Expression and localization of <t>S100A14</t> and S100A16 in human breast cancer cell lines. A , Relative mRNA expression levels measured by real-time <t>RT-PCR.</t> Expression levels were normalized to β-actin levels within the same sample. B , Protein expression of S100A14 and S100A16. Expression levels and subcellular localization were visualized using immunofluorescence staining. Scale bar; 50 μm. C , Subcellular localization of the S100A14 protein on the cell membrane. Z-axis images of confluent MCF7 cells were constructed using confocal laser scanning microscopy. Scale bar; 10 μm. D , Effect of omission of cell permeabilization on the immunofluorescent staining of S100A14. Following fixation with 4% paraformaldehyde, the cells were treated with or without 0.1% Triton X-100 in PBS prior to staining. Scale bar; 50 μm. E , Ca 2 -independent localization of S100A14 on the cell membrane. MCF7 cells transfected with the S100A14-GFP expression vector were observed by fluorescence microscopy over 180 min after addition of 10 mM EGTA in PBS. Scale bar; 50 μm.
    Pcr Amplification, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 21524 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr amplification/product/TaKaRa
    Average 99 stars, based on 21524 article reviews
    Price from $9.99 to $1999.99
    pcr amplification - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa sybr premix ex taq
    Expression and localization of <t>S100A14</t> and S100A16 in human breast cancer cell lines. A , Relative mRNA expression levels measured by real-time <t>RT-PCR.</t> Expression levels were normalized to β-actin levels within the same sample. B , Protein expression of S100A14 and S100A16. Expression levels and subcellular localization were visualized using immunofluorescence staining. Scale bar; 50 μm. C , Subcellular localization of the S100A14 protein on the cell membrane. Z-axis images of confluent MCF7 cells were constructed using confocal laser scanning microscopy. Scale bar; 10 μm. D , Effect of omission of cell permeabilization on the immunofluorescent staining of S100A14. Following fixation with 4% paraformaldehyde, the cells were treated with or without 0.1% Triton X-100 in PBS prior to staining. Scale bar; 50 μm. E , Ca 2 -independent localization of S100A14 on the cell membrane. MCF7 cells transfected with the S100A14-GFP expression vector were observed by fluorescence microscopy over 180 min after addition of 10 mM EGTA in PBS. Scale bar; 50 μm.
    Sybr Premix Ex Taq, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 49731 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr premix ex taq/product/TaKaRa
    Average 99 stars, based on 49731 article reviews
    Price from $9.99 to $1999.99
    sybr premix ex taq - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa primescript rt reagent kit
    Expression and localization of <t>S100A14</t> and S100A16 in human breast cancer cell lines. A , Relative mRNA expression levels measured by real-time <t>RT-PCR.</t> Expression levels were normalized to β-actin levels within the same sample. B , Protein expression of S100A14 and S100A16. Expression levels and subcellular localization were visualized using immunofluorescence staining. Scale bar; 50 μm. C , Subcellular localization of the S100A14 protein on the cell membrane. Z-axis images of confluent MCF7 cells were constructed using confocal laser scanning microscopy. Scale bar; 10 μm. D , Effect of omission of cell permeabilization on the immunofluorescent staining of S100A14. Following fixation with 4% paraformaldehyde, the cells were treated with or without 0.1% Triton X-100 in PBS prior to staining. Scale bar; 50 μm. E , Ca 2 -independent localization of S100A14 on the cell membrane. MCF7 cells transfected with the S100A14-GFP expression vector were observed by fluorescence microscopy over 180 min after addition of 10 mM EGTA in PBS. Scale bar; 50 μm.
    Primescript Rt Reagent Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 72258 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primescript rt reagent kit/product/TaKaRa
    Average 99 stars, based on 72258 article reviews
    Price from $9.99 to $1999.99
    primescript rt reagent kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    95
    Thermo Fisher revertaid first strand cdna synthesis kit
    Expression and localization of <t>S100A14</t> and S100A16 in human breast cancer cell lines. A , Relative mRNA expression levels measured by real-time <t>RT-PCR.</t> Expression levels were normalized to β-actin levels within the same sample. B , Protein expression of S100A14 and S100A16. Expression levels and subcellular localization were visualized using immunofluorescence staining. Scale bar; 50 μm. C , Subcellular localization of the S100A14 protein on the cell membrane. Z-axis images of confluent MCF7 cells were constructed using confocal laser scanning microscopy. Scale bar; 10 μm. D , Effect of omission of cell permeabilization on the immunofluorescent staining of S100A14. Following fixation with 4% paraformaldehyde, the cells were treated with or without 0.1% Triton X-100 in PBS prior to staining. Scale bar; 50 μm. E , Ca 2 -independent localization of S100A14 on the cell membrane. MCF7 cells transfected with the S100A14-GFP expression vector were observed by fluorescence microscopy over 180 min after addition of 10 mM EGTA in PBS. Scale bar; 50 μm.
    Revertaid First Strand Cdna Synthesis Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 40122 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/revertaid first strand cdna synthesis kit/product/Thermo Fisher
    Average 95 stars, based on 40122 article reviews
    Price from $9.99 to $1999.99
    revertaid first strand cdna synthesis kit - by Bioz Stars, 2020-09
    95/100 stars
      Buy from Supplier

    99
    TaKaRa primescript rt pcr kit
    2.8. <t>SYBR</t> green real-time reverse transcription-polymerase chain reaction <t>(RT-PCR)</t>
    Primescript Rt Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 5027 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primescript rt pcr kit/product/TaKaRa
    Average 99 stars, based on 5027 article reviews
    Price from $9.99 to $1999.99
    primescript rt pcr kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa ex taq polymerase
    Methylation profiles of spermatogenic cell-specific intronless genes are classified three groups in juvenile mouse testis. COBRA was carried out to examine the methylation status of the selected genes, the CpG island in the Oaz1 gene, and constitutively methylated endogenous IAP retroviruses in genomic DNA isolated from D7, D14, D21, and D35 testes and adult liver. Sodium bisulfite-treated DNA was amplified with specific primers (details in Table S3 ) and digested with HpyCH4 IV (recognition site: ACGT), <t>Taq</t> I (TCGA) or Acc II (CGCG).
    Ex Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 7747 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ex taq polymerase/product/TaKaRa
    Average 99 stars, based on 7747 article reviews
    Price from $9.99 to $1999.99
    ex taq polymerase - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa m mlv reverse transcriptase
    (A) Laser-scanning confocal microscope was used to detect the distribution of ZIP4 in HepG2, BEL7402, and HL-7702 cells (magnification: ×1260). Cells were incubated in a cell culture dish, fixed with paraformaldehyde for 30 min, and permeabilized with 0.1% Triton X-100 for 5 min at room temperature and incubated with primary polyclonal rabbit anti-human ZIP4 antibody. Cells were then washed and incubated with Alexa Fluor 488 donkey anti-rabbit IgG. The green signal represents staining for ZIP4 protein. ZIP4 located in cell membranes of HL-7702 cells, but not in nuclei. ZIP4 accumulated in the nuclei in HepG2 and BEL7402 cells. (B) The mRNA levels of ZIP4 in seven liver cell lines. We used the <t>TRIzol</t> reagent (Invitrogen) to extract total RNA and performed cDNA synthesis using <t>M-MLV</t> reverse transcriptase (TaKaRa, Dalian, China). ß-actin was the endogenous control. All the samples were normalized to human ß-actin according to the 2 -ΔΔCT method. ZIP4 mRNA levels in immortalized liver cells (HL-7702) was the negative control. The x- axis represents multiples of mRNA levels in HL-7702 cells. (C) ZIP4 protein levels in eight liver cell lines. (D) The mRNA levels of ZIP4 in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. β-actin was used as an internal control. (E) ZIP4 protein levels in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. (F) ZIP4 mRNA levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and negative control (NEG). β-actin was used as an internal control. (G) ZIP4 protein levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and a negative control (NEG). Data are from one of three repeated independent experiments. * P
    M Mlv Reverse Transcriptase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 4278 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m mlv reverse transcriptase/product/TaKaRa
    Average 99 stars, based on 4278 article reviews
    Price from $9.99 to $1999.99
    m mlv reverse transcriptase - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher steponeplus real time pcr system
    (A) Laser-scanning confocal microscope was used to detect the distribution of ZIP4 in HepG2, BEL7402, and HL-7702 cells (magnification: ×1260). Cells were incubated in a cell culture dish, fixed with paraformaldehyde for 30 min, and permeabilized with 0.1% Triton X-100 for 5 min at room temperature and incubated with primary polyclonal rabbit anti-human ZIP4 antibody. Cells were then washed and incubated with Alexa Fluor 488 donkey anti-rabbit IgG. The green signal represents staining for ZIP4 protein. ZIP4 located in cell membranes of HL-7702 cells, but not in nuclei. ZIP4 accumulated in the nuclei in HepG2 and BEL7402 cells. (B) The mRNA levels of ZIP4 in seven liver cell lines. We used the <t>TRIzol</t> reagent (Invitrogen) to extract total RNA and performed cDNA synthesis using <t>M-MLV</t> reverse transcriptase (TaKaRa, Dalian, China). ß-actin was the endogenous control. All the samples were normalized to human ß-actin according to the 2 -ΔΔCT method. ZIP4 mRNA levels in immortalized liver cells (HL-7702) was the negative control. The x- axis represents multiples of mRNA levels in HL-7702 cells. (C) ZIP4 protein levels in eight liver cell lines. (D) The mRNA levels of ZIP4 in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. β-actin was used as an internal control. (E) ZIP4 protein levels in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. (F) ZIP4 mRNA levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and negative control (NEG). β-actin was used as an internal control. (G) ZIP4 protein levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and a negative control (NEG). Data are from one of three repeated independent experiments. * P
    Steponeplus Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 52235 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/steponeplus real time pcr system/product/Thermo Fisher
    Average 99 stars, based on 52235 article reviews
    Price from $9.99 to $1999.99
    steponeplus real time pcr system - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa primescript first strand cdna synthesis kit
    (A) Laser-scanning confocal microscope was used to detect the distribution of ZIP4 in HepG2, BEL7402, and HL-7702 cells (magnification: ×1260). Cells were incubated in a cell culture dish, fixed with paraformaldehyde for 30 min, and permeabilized with 0.1% Triton X-100 for 5 min at room temperature and incubated with primary polyclonal rabbit anti-human ZIP4 antibody. Cells were then washed and incubated with Alexa Fluor 488 donkey anti-rabbit IgG. The green signal represents staining for ZIP4 protein. ZIP4 located in cell membranes of HL-7702 cells, but not in nuclei. ZIP4 accumulated in the nuclei in HepG2 and BEL7402 cells. (B) The mRNA levels of ZIP4 in seven liver cell lines. We used the <t>TRIzol</t> reagent (Invitrogen) to extract total RNA and performed cDNA synthesis using <t>M-MLV</t> reverse transcriptase (TaKaRa, Dalian, China). ß-actin was the endogenous control. All the samples were normalized to human ß-actin according to the 2 -ΔΔCT method. ZIP4 mRNA levels in immortalized liver cells (HL-7702) was the negative control. The x- axis represents multiples of mRNA levels in HL-7702 cells. (C) ZIP4 protein levels in eight liver cell lines. (D) The mRNA levels of ZIP4 in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. β-actin was used as an internal control. (E) ZIP4 protein levels in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. (F) ZIP4 mRNA levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and negative control (NEG). β-actin was used as an internal control. (G) ZIP4 protein levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and a negative control (NEG). Data are from one of three repeated independent experiments. * P
    Primescript First Strand Cdna Synthesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 2734 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primescript first strand cdna synthesis kit/product/TaKaRa
    Average 99 stars, based on 2734 article reviews
    Price from $9.99 to $1999.99
    primescript first strand cdna synthesis kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa amv reverse transcriptase
    ( A ) Sequences of the 25 nt template ( III ) and 20 nt primer ( IV ). ( B ) PAGE of extension reactions beyond the 1-deaza-dG residue in the template. ( C ) PAGE of extension reactions subsequent to 1-deaza-dG in the primer with Therminator™ DNA polymerase and ( D ) with <t>AMV</t> reverse transcriptase. Conditions used are described in Materials and Methods. Added <t>dNTPs</t> are indicated at the top of each gel. Z indicates 1-deaza-dGTP; P indicates reaction without dNTP.
    Amv Reverse Transcriptase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 3369 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/amv reverse transcriptase/product/TaKaRa
    Average 99 stars, based on 3369 article reviews
    Price from $9.99 to $1999.99
    amv reverse transcriptase - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher high capacity cdna reverse transcription kit
    Cloning of EXA1 homologs in Nicotiana benthamiana and their functional analysis in potexvirus infection. ( a ) Schematic image of the <t>cDNA</t> structure of NbEXA1a . GYF domain- and eIF4E-binding motif-encoding regions are depicted by stripes. Target regions for virus-induced gene silencing (VIGS) and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) are indicated by bars under the image. ( b ) Morphological phenotypes of NbEXA1 -silenced and control plants. Photographs were taken from the top (upper) and side (bottom) of plants at 27 days post-inoculation (dpi). Bars = 5 cm. ( c ) Relative accumulation of NbEXA1 mRNA in NbEXA1 -silenced and control plants. Total <t>RNA</t> was extracted at 27 dpi and analyzed using qRT-PCR. The mean level of NbEXA1 transcript in control plants was used as the standard (1.0), and that in NbEXA1 -silenced plants is shown above the bar. Error bars indicate standard deviation (SD) of 10 samples. ** P
    High Capacity Cdna Reverse Transcription Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 117891 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/high capacity cdna reverse transcription kit/product/Thermo Fisher
    Average 99 stars, based on 117891 article reviews
    Price from $9.99 to $1999.99
    high capacity cdna reverse transcription kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa primescript 1st strand cdna synthesis kit
    Cloning of EXA1 homologs in Nicotiana benthamiana and their functional analysis in potexvirus infection. ( a ) Schematic image of the <t>cDNA</t> structure of NbEXA1a . GYF domain- and eIF4E-binding motif-encoding regions are depicted by stripes. Target regions for virus-induced gene silencing (VIGS) and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) are indicated by bars under the image. ( b ) Morphological phenotypes of NbEXA1 -silenced and control plants. Photographs were taken from the top (upper) and side (bottom) of plants at 27 days post-inoculation (dpi). Bars = 5 cm. ( c ) Relative accumulation of NbEXA1 mRNA in NbEXA1 -silenced and control plants. Total <t>RNA</t> was extracted at 27 dpi and analyzed using qRT-PCR. The mean level of NbEXA1 transcript in control plants was used as the standard (1.0), and that in NbEXA1 -silenced plants is shown above the bar. Error bars indicate standard deviation (SD) of 10 samples. ** P
    Primescript 1st Strand Cdna Synthesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 7752 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primescript 1st strand cdna synthesis kit/product/TaKaRa
    Average 99 stars, based on 7752 article reviews
    Price from $9.99 to $1999.99
    primescript 1st strand cdna synthesis kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa smart seq v4 ultra low input rna kit
    Comparison of FL-cDNA-Seq and Illumina <t>RNA-Seq.</t> (A) The gene expression of FL-cDNA-Seq of LC2/ad (R9.4) was compared with that of TruSeq RNA (left) and <t>SMART-Seq</t> (right). Pearson correlation coefficients are shown on the graph. (B) Influence of sequencing depth on the estimation of gene expression level and gene detection. Reads for each method were randomly sampled in triplicate. The average of the Pearson correlation coefficients between TruSeq RNA and randomly sampled data for FL-cDNA-Seq and SMART-Seq is shown (left). The average number of genes with an expression level of more than 1 tpm or ppm is shown (right). (C) Comparison to qRT-qPCR. Forty-four genes detected by all methods were analyzed. The gene expression of these genes was normalized to GAPDH. Pearson correlation coefficients are shown on the graph. qRT-qPCR data of LC2/ad was obtained as in our previous study. 16
    Smart Seq V4 Ultra Low Input Rna Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1758 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/smart seq v4 ultra low input rna kit/product/TaKaRa
    Average 99 stars, based on 1758 article reviews
    Price from $9.99 to $1999.99
    smart seq v4 ultra low input rna kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa dna polymerase
    Detection and determination of Has2 gene mutation A 454 bp region within exon2 of rat Has2 that contains the Cas9 mutation site was <t>PCR</t> amplified from genomic <t>DNA</t> of each cell clone as a template. The amplified product was gel-purified and then digested with Pas1. The final products were analyzed by agarose gel electrophoresis. DNA standards (std) are labeled. Panel A. Products amplified from RCS-o WT cells and clones 1 thru 7. Panel B. Pas1 digestion products of the amplified 454 bp products shown in panel A. Panel C. Products amplified from RCS-Cas9 WT cells and clones 3, 7, 39, 43, 53, 80 and 88. Panel D. Pas1 digestion products of the amplified 454 bp products shown in panel C. Panel E. Alignment of the targeted rat Has2 genomic sequences from RCS-Cas9 clone #7, allele 1 and allele 2 with the WT RCS-Cas9 cells. Deleted bases are indicated.
    Dna Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 2424 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna polymerase/product/TaKaRa
    Average 99 stars, based on 2424 article reviews
    Price from $9.99 to $1999.99
    dna polymerase - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa advantage 2 polymerase mix
    Detection and determination of Has2 gene mutation A 454 bp region within exon2 of rat Has2 that contains the Cas9 mutation site was <t>PCR</t> amplified from genomic <t>DNA</t> of each cell clone as a template. The amplified product was gel-purified and then digested with Pas1. The final products were analyzed by agarose gel electrophoresis. DNA standards (std) are labeled. Panel A. Products amplified from RCS-o WT cells and clones 1 thru 7. Panel B. Pas1 digestion products of the amplified 454 bp products shown in panel A. Panel C. Products amplified from RCS-Cas9 WT cells and clones 3, 7, 39, 43, 53, 80 and 88. Panel D. Pas1 digestion products of the amplified 454 bp products shown in panel C. Panel E. Alignment of the targeted rat Has2 genomic sequences from RCS-Cas9 clone #7, allele 1 and allele 2 with the WT RCS-Cas9 cells. Deleted bases are indicated.
    Advantage 2 Polymerase Mix, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 2759 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/advantage 2 polymerase mix/product/TaKaRa
    Average 99 stars, based on 2759 article reviews
    Price from $9.99 to $1999.99
    advantage 2 polymerase mix - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa advantage 2 pcr kit
    Detection and determination of Has2 gene mutation A 454 bp region within exon2 of rat Has2 that contains the Cas9 mutation site was <t>PCR</t> amplified from genomic <t>DNA</t> of each cell clone as a template. The amplified product was gel-purified and then digested with Pas1. The final products were analyzed by agarose gel electrophoresis. DNA standards (std) are labeled. Panel A. Products amplified from RCS-o WT cells and clones 1 thru 7. Panel B. Pas1 digestion products of the amplified 454 bp products shown in panel A. Panel C. Products amplified from RCS-Cas9 WT cells and clones 3, 7, 39, 43, 53, 80 and 88. Panel D. Pas1 digestion products of the amplified 454 bp products shown in panel C. Panel E. Alignment of the targeted rat Has2 genomic sequences from RCS-Cas9 clone #7, allele 1 and allele 2 with the WT RCS-Cas9 cells. Deleted bases are indicated.
    Advantage 2 Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 3881 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/advantage 2 pcr kit/product/TaKaRa
    Average 99 stars, based on 3881 article reviews
    Price from $9.99 to $1999.99
    advantage 2 pcr kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher power sybr green pcr master mix
    Detection and determination of Has2 gene mutation A 454 bp region within exon2 of rat Has2 that contains the Cas9 mutation site was <t>PCR</t> amplified from genomic <t>DNA</t> of each cell clone as a template. The amplified product was gel-purified and then digested with Pas1. The final products were analyzed by agarose gel electrophoresis. DNA standards (std) are labeled. Panel A. Products amplified from RCS-o WT cells and clones 1 thru 7. Panel B. Pas1 digestion products of the amplified 454 bp products shown in panel A. Panel C. Products amplified from RCS-Cas9 WT cells and clones 3, 7, 39, 43, 53, 80 and 88. Panel D. Pas1 digestion products of the amplified 454 bp products shown in panel C. Panel E. Alignment of the targeted rat Has2 genomic sequences from RCS-Cas9 clone #7, allele 1 and allele 2 with the WT RCS-Cas9 cells. Deleted bases are indicated.
    Power Sybr Green Pcr Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 69800 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/power sybr green pcr master mix/product/Thermo Fisher
    Average 99 stars, based on 69800 article reviews
    Price from $9.99 to $1999.99
    power sybr green pcr master mix - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa gdna eraser
    Screening of PAF-AH knockout <t>transformants.</t> (A) Physical map of homologous recombination. (B) Identification of transformants by PCR: 1, fragment of the hygromycin B resistance gene ( hyg ); 2, amplified fragment of PAF-AH from genomic DNA; 3, amplified T-DNA fragments adjusted to the right border of plasmid; 4, fragment amplified with primers PAF-AH-UP-F and Hyg-R. (C) Southern blot to verify T-DNA insertion copies of knockout transformants. The 450 bp fragment amplified from the hyg gene was labeled with 32 P-dCTP as probe 1. (D) Southern blot to confirm PAF-AH was replaced using the 500 bp fragment of PAF-AH as probe 2. WT, wild type strain; P, plasmid; KO15 and KO40: PAF-AH knockout transformants; M: DNA marker.
    Gdna Eraser, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 27214 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gdna eraser/product/TaKaRa
    Average 99 stars, based on 27214 article reviews
    Price from $9.99 to $1999.99
    gdna eraser - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    TaKaRa la taq polymerase
    Comparison of the <t>BmP5CR1</t> genomic region between Daizo (+ Lg /+ Lg ) and DH6 ( Lg / Lg ). ( a ) Genomic PCR analysis of the three parts of BmP5CR1 by Ex or LA <t>Taq</t> polymerase HS. M is the DNA ladder marker. ( b ) Schematic representation of the BmP5CR1 gene structure. Lines and rectangular boxes show introns and exons, respectively. White boxes indicate the untranslated regions (UTRs), and grey boxes are the coding regions. Arrows indicate differences between Daizo and DH6. Arrowheads are primer sites used in this study. Numbers show the length (bp) of each region
    La Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 5601 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/la taq polymerase/product/TaKaRa
    Average 99 stars, based on 5601 article reviews
    Price from $9.99 to $1999.99
    la taq polymerase - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher stepone real time pcr system
    Comparison of the <t>BmP5CR1</t> genomic region between Daizo (+ Lg /+ Lg ) and DH6 ( Lg / Lg ). ( a ) Genomic PCR analysis of the three parts of BmP5CR1 by Ex or LA <t>Taq</t> polymerase HS. M is the DNA ladder marker. ( b ) Schematic representation of the BmP5CR1 gene structure. Lines and rectangular boxes show introns and exons, respectively. White boxes indicate the untranslated regions (UTRs), and grey boxes are the coding regions. Arrows indicate differences between Daizo and DH6. Arrowheads are primer sites used in this study. Numbers show the length (bp) of each region
    Stepone Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 19014 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stepone real time pcr system/product/Thermo Fisher
    Average 99 stars, based on 19014 article reviews
    Price from $9.99 to $1999.99
    stepone real time pcr system - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Quantification of the mRNA transcriptional levels of EGFP (A) and IL2 (B). Tumor cells in 6-well plate were infected with the indicated rNDV-EGFPs or rNDV-IL2s at MOI of 10. Total RNA in infected cells was extracted using Trizol reagent at 48 hpi, and qRT-PCR was performed to quantify mRNA transcriptional levels of EGFP and IL2, respectively. Transcriptional levels of EGFP and IL2 genes are calculated relative to the β-actin, and the data are expressed as fold changes with SEM (* P

    Journal: PLoS ONE

    Article Title: Identification of Optimal Insertion Site in Recombinant Newcastle Disease Virus (rNDV) Vector Expressing Foreign Gene to Enhance Its Anti-Tumor Effect

    doi: 10.1371/journal.pone.0164723

    Figure Lengend Snippet: Quantification of the mRNA transcriptional levels of EGFP (A) and IL2 (B). Tumor cells in 6-well plate were infected with the indicated rNDV-EGFPs or rNDV-IL2s at MOI of 10. Total RNA in infected cells was extracted using Trizol reagent at 48 hpi, and qRT-PCR was performed to quantify mRNA transcriptional levels of EGFP and IL2, respectively. Transcriptional levels of EGFP and IL2 genes are calculated relative to the β-actin, and the data are expressed as fold changes with SEM (* P

    Article Snippet: At the same time, total RNA in infected cells was extracted using Trizol reagent, and quantitative real-time PCR (qRT-PCR) was used to quantify mRNA levels of EGFP and IL2 transcribed from rNDV-EGFPs and rNDV-IL2s using the GeneAmp kit (Applied Biosystems; ABI) [ ].

    Techniques: Infection, Quantitative RT-PCR

    Expression and localization of S100A14 and S100A16 in human breast cancer cell lines. A , Relative mRNA expression levels measured by real-time RT-PCR. Expression levels were normalized to β-actin levels within the same sample. B , Protein expression of S100A14 and S100A16. Expression levels and subcellular localization were visualized using immunofluorescence staining. Scale bar; 50 μm. C , Subcellular localization of the S100A14 protein on the cell membrane. Z-axis images of confluent MCF7 cells were constructed using confocal laser scanning microscopy. Scale bar; 10 μm. D , Effect of omission of cell permeabilization on the immunofluorescent staining of S100A14. Following fixation with 4% paraformaldehyde, the cells were treated with or without 0.1% Triton X-100 in PBS prior to staining. Scale bar; 50 μm. E , Ca 2 -independent localization of S100A14 on the cell membrane. MCF7 cells transfected with the S100A14-GFP expression vector were observed by fluorescence microscopy over 180 min after addition of 10 mM EGTA in PBS. Scale bar; 50 μm.

    Journal: BMC Cancer

    Article Title: Co-expression of S100A14 and S100A16 correlates with a poor prognosis in human breast cancer and promotes cancer cell invasion

    doi: 10.1186/s12885-015-1059-6

    Figure Lengend Snippet: Expression and localization of S100A14 and S100A16 in human breast cancer cell lines. A , Relative mRNA expression levels measured by real-time RT-PCR. Expression levels were normalized to β-actin levels within the same sample. B , Protein expression of S100A14 and S100A16. Expression levels and subcellular localization were visualized using immunofluorescence staining. Scale bar; 50 μm. C , Subcellular localization of the S100A14 protein on the cell membrane. Z-axis images of confluent MCF7 cells were constructed using confocal laser scanning microscopy. Scale bar; 10 μm. D , Effect of omission of cell permeabilization on the immunofluorescent staining of S100A14. Following fixation with 4% paraformaldehyde, the cells were treated with or without 0.1% Triton X-100 in PBS prior to staining. Scale bar; 50 μm. E , Ca 2 -independent localization of S100A14 on the cell membrane. MCF7 cells transfected with the S100A14-GFP expression vector were observed by fluorescence microscopy over 180 min after addition of 10 mM EGTA in PBS. Scale bar; 50 μm.

    Article Snippet: Construction of S100A14 and S100A16 expression vectors and transfection cDNA expression vectors for the human S100A14 and S100A16 genes were constructed by PCR amplification of their coding regions using cDNAs derived from MCF7 cells as templates and specific primers, followed by cloning of the genes into a pEGFP expression vector (Takara-Clontech, Shiga, Japan).

    Techniques: Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Construct, Confocal Laser Scanning Microscopy, Transfection, Plasmid Preparation, Fluorescence, Microscopy

    2.8. SYBR green real-time reverse transcription-polymerase chain reaction (RT-PCR)

    Journal: Journal of Zhejiang University. Science. B

    Article Title: Growth factors induce the improved cardiac remodeling in autologous mesenchymal stem cell-implanted failing rat hearts

    doi: 10.1631/jzus.B0900244

    Figure Lengend Snippet: 2.8. SYBR green real-time reverse transcription-polymerase chain reaction (RT-PCR)

    Article Snippet: The SYBR green real-time RT-PCR amplifications were performed using SYBR® PrimeScript™ RT-PCR Kit (TaKaRa Biotech Co.) and Applied Biosystems 7500 System (Applied Biosystems, USA).

    Techniques: SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction

    Methylation profiles of spermatogenic cell-specific intronless genes are classified three groups in juvenile mouse testis. COBRA was carried out to examine the methylation status of the selected genes, the CpG island in the Oaz1 gene, and constitutively methylated endogenous IAP retroviruses in genomic DNA isolated from D7, D14, D21, and D35 testes and adult liver. Sodium bisulfite-treated DNA was amplified with specific primers (details in Table S3 ) and digested with HpyCH4 IV (recognition site: ACGT), Taq I (TCGA) or Acc II (CGCG).

    Journal: PLoS ONE

    Article Title: Distinct DNA Methylation Dynamics of Spermatogenic Cell-Specific Intronless Genes Is Associated with CpG Content

    doi: 10.1371/journal.pone.0043658

    Figure Lengend Snippet: Methylation profiles of spermatogenic cell-specific intronless genes are classified three groups in juvenile mouse testis. COBRA was carried out to examine the methylation status of the selected genes, the CpG island in the Oaz1 gene, and constitutively methylated endogenous IAP retroviruses in genomic DNA isolated from D7, D14, D21, and D35 testes and adult liver. Sodium bisulfite-treated DNA was amplified with specific primers (details in Table S3 ) and digested with HpyCH4 IV (recognition site: ACGT), Taq I (TCGA) or Acc II (CGCG).

    Article Snippet: Reverse Transcription (RT)-PCR One microgram of total RNA was treated with DNase I at 37°C for 20 min, and the cDNA was reverse-transcribed using SuperScript III (Invitrogen) at 50°C for 60 min. Ex Taq Polymerase (TaKaRa) was then used for PCR with the primers and conditions shown in .

    Techniques: Methylation, Combined Bisulfite Restriction Analysis Assay, Isolation, Amplification

    (A) Laser-scanning confocal microscope was used to detect the distribution of ZIP4 in HepG2, BEL7402, and HL-7702 cells (magnification: ×1260). Cells were incubated in a cell culture dish, fixed with paraformaldehyde for 30 min, and permeabilized with 0.1% Triton X-100 for 5 min at room temperature and incubated with primary polyclonal rabbit anti-human ZIP4 antibody. Cells were then washed and incubated with Alexa Fluor 488 donkey anti-rabbit IgG. The green signal represents staining for ZIP4 protein. ZIP4 located in cell membranes of HL-7702 cells, but not in nuclei. ZIP4 accumulated in the nuclei in HepG2 and BEL7402 cells. (B) The mRNA levels of ZIP4 in seven liver cell lines. We used the TRIzol reagent (Invitrogen) to extract total RNA and performed cDNA synthesis using M-MLV reverse transcriptase (TaKaRa, Dalian, China). ß-actin was the endogenous control. All the samples were normalized to human ß-actin according to the 2 -ΔΔCT method. ZIP4 mRNA levels in immortalized liver cells (HL-7702) was the negative control. The x- axis represents multiples of mRNA levels in HL-7702 cells. (C) ZIP4 protein levels in eight liver cell lines. (D) The mRNA levels of ZIP4 in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. β-actin was used as an internal control. (E) ZIP4 protein levels in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. (F) ZIP4 mRNA levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and negative control (NEG). β-actin was used as an internal control. (G) ZIP4 protein levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and a negative control (NEG). Data are from one of three repeated independent experiments. * P

    Journal: International Journal of Biological Sciences

    Article Title: ZIP4, a Novel Determinant of Tumor Invasion in Hepatocellular Carcinoma, Contributes to Tumor Recurrence after Liver Transplantation

    doi: 10.7150/ijbs.7401

    Figure Lengend Snippet: (A) Laser-scanning confocal microscope was used to detect the distribution of ZIP4 in HepG2, BEL7402, and HL-7702 cells (magnification: ×1260). Cells were incubated in a cell culture dish, fixed with paraformaldehyde for 30 min, and permeabilized with 0.1% Triton X-100 for 5 min at room temperature and incubated with primary polyclonal rabbit anti-human ZIP4 antibody. Cells were then washed and incubated with Alexa Fluor 488 donkey anti-rabbit IgG. The green signal represents staining for ZIP4 protein. ZIP4 located in cell membranes of HL-7702 cells, but not in nuclei. ZIP4 accumulated in the nuclei in HepG2 and BEL7402 cells. (B) The mRNA levels of ZIP4 in seven liver cell lines. We used the TRIzol reagent (Invitrogen) to extract total RNA and performed cDNA synthesis using M-MLV reverse transcriptase (TaKaRa, Dalian, China). ß-actin was the endogenous control. All the samples were normalized to human ß-actin according to the 2 -ΔΔCT method. ZIP4 mRNA levels in immortalized liver cells (HL-7702) was the negative control. The x- axis represents multiples of mRNA levels in HL-7702 cells. (C) ZIP4 protein levels in eight liver cell lines. (D) The mRNA levels of ZIP4 in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. β-actin was used as an internal control. (E) ZIP4 protein levels in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. (F) ZIP4 mRNA levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and negative control (NEG). β-actin was used as an internal control. (G) ZIP4 protein levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and a negative control (NEG). Data are from one of three repeated independent experiments. * P

    Article Snippet: Real-time polymerase chain reaction RNA was extracted using the TRIzol reagent (Invitrogen) and used for cDNA synthesis with a M-MLV Reverse Transcriptase (TaKaRa, Dalian, China).

    Techniques: Microscopy, Incubation, Cell Culture, Staining, Negative Control, shRNA, Over Expression

    ( A ) Sequences of the 25 nt template ( III ) and 20 nt primer ( IV ). ( B ) PAGE of extension reactions beyond the 1-deaza-dG residue in the template. ( C ) PAGE of extension reactions subsequent to 1-deaza-dG in the primer with Therminator™ DNA polymerase and ( D ) with AMV reverse transcriptase. Conditions used are described in Materials and Methods. Added dNTPs are indicated at the top of each gel. Z indicates 1-deaza-dGTP; P indicates reaction without dNTP.

    Journal: Nucleic Acids Research

    Article Title: A new, but old, nucleoside analog: the first synthesis of 1-deaza-2?-deoxyguanosine and its properties as a nucleoside and as oligodeoxynucleotides

    doi: 10.1093/nar/gkh154

    Figure Lengend Snippet: ( A ) Sequences of the 25 nt template ( III ) and 20 nt primer ( IV ). ( B ) PAGE of extension reactions beyond the 1-deaza-dG residue in the template. ( C ) PAGE of extension reactions subsequent to 1-deaza-dG in the primer with Therminator™ DNA polymerase and ( D ) with AMV reverse transcriptase. Conditions used are described in Materials and Methods. Added dNTPs are indicated at the top of each gel. Z indicates 1-deaza-dGTP; P indicates reaction without dNTP.

    Article Snippet: AMV reverse transcriptase, M-MLV reverse transcriptase and dNTPs were purchased from Takara Biomedicals (Japan).

    Techniques: Polyacrylamide Gel Electrophoresis

    PAGE of single nucleotide insertion studies ( A ) with Therminator™ DNA polymerase (reaction mixtures being incubated at 74°C for 10 min), ( B ) with AMV reverse transcriptase (reaction mixtures being incubated at 37°C for 30 min) and ( C ) with M-MLV reverse transcriptase (reactions mixture being incubated at 37°C for 30 min). Conditions used are described in Materials and Methods. Both X in the template sequence and added dNTP are indicated at the top of each gel. Z indicates 1-deaza-dGTP; P indicates reaction without dNTP.

    Journal: Nucleic Acids Research

    Article Title: A new, but old, nucleoside analog: the first synthesis of 1-deaza-2?-deoxyguanosine and its properties as a nucleoside and as oligodeoxynucleotides

    doi: 10.1093/nar/gkh154

    Figure Lengend Snippet: PAGE of single nucleotide insertion studies ( A ) with Therminator™ DNA polymerase (reaction mixtures being incubated at 74°C for 10 min), ( B ) with AMV reverse transcriptase (reaction mixtures being incubated at 37°C for 30 min) and ( C ) with M-MLV reverse transcriptase (reactions mixture being incubated at 37°C for 30 min). Conditions used are described in Materials and Methods. Both X in the template sequence and added dNTP are indicated at the top of each gel. Z indicates 1-deaza-dGTP; P indicates reaction without dNTP.

    Article Snippet: AMV reverse transcriptase, M-MLV reverse transcriptase and dNTPs were purchased from Takara Biomedicals (Japan).

    Techniques: Polyacrylamide Gel Electrophoresis, Incubation, Sequencing

    Cloning of EXA1 homologs in Nicotiana benthamiana and their functional analysis in potexvirus infection. ( a ) Schematic image of the cDNA structure of NbEXA1a . GYF domain- and eIF4E-binding motif-encoding regions are depicted by stripes. Target regions for virus-induced gene silencing (VIGS) and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) are indicated by bars under the image. ( b ) Morphological phenotypes of NbEXA1 -silenced and control plants. Photographs were taken from the top (upper) and side (bottom) of plants at 27 days post-inoculation (dpi). Bars = 5 cm. ( c ) Relative accumulation of NbEXA1 mRNA in NbEXA1 -silenced and control plants. Total RNA was extracted at 27 dpi and analyzed using qRT-PCR. The mean level of NbEXA1 transcript in control plants was used as the standard (1.0), and that in NbEXA1 -silenced plants is shown above the bar. Error bars indicate standard deviation (SD) of 10 samples. ** P

    Journal: Scientific Reports

    Article Title: Functional conservation of EXA1 among diverse plant species for the infection by a family of plant viruses

    doi: 10.1038/s41598-019-42400-w

    Figure Lengend Snippet: Cloning of EXA1 homologs in Nicotiana benthamiana and their functional analysis in potexvirus infection. ( a ) Schematic image of the cDNA structure of NbEXA1a . GYF domain- and eIF4E-binding motif-encoding regions are depicted by stripes. Target regions for virus-induced gene silencing (VIGS) and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) are indicated by bars under the image. ( b ) Morphological phenotypes of NbEXA1 -silenced and control plants. Photographs were taken from the top (upper) and side (bottom) of plants at 27 days post-inoculation (dpi). Bars = 5 cm. ( c ) Relative accumulation of NbEXA1 mRNA in NbEXA1 -silenced and control plants. Total RNA was extracted at 27 dpi and analyzed using qRT-PCR. The mean level of NbEXA1 transcript in control plants was used as the standard (1.0), and that in NbEXA1 -silenced plants is shown above the bar. Error bars indicate standard deviation (SD) of 10 samples. ** P

    Article Snippet: RNA isolation, RT-PCR, and qRT-PCR Total RNA was extracted from the plants using ISOGEN (Nippon Gene) and treated with DNase I (Takara Bio, Shiga, Japan), or using the ISOSPIN Plant RNA Kit (Nippongene) following the manufacturer’s instructions. cDNA was synthesized using a High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Clone Assay, Functional Assay, Infection, Binding Assay, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Standard Deviation

    Comparison of FL-cDNA-Seq and Illumina RNA-Seq. (A) The gene expression of FL-cDNA-Seq of LC2/ad (R9.4) was compared with that of TruSeq RNA (left) and SMART-Seq (right). Pearson correlation coefficients are shown on the graph. (B) Influence of sequencing depth on the estimation of gene expression level and gene detection. Reads for each method were randomly sampled in triplicate. The average of the Pearson correlation coefficients between TruSeq RNA and randomly sampled data for FL-cDNA-Seq and SMART-Seq is shown (left). The average number of genes with an expression level of more than 1 tpm or ppm is shown (right). (C) Comparison to qRT-qPCR. Forty-four genes detected by all methods were analyzed. The gene expression of these genes was normalized to GAPDH. Pearson correlation coefficients are shown on the graph. qRT-qPCR data of LC2/ad was obtained as in our previous study. 16

    Journal: DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes

    Article Title: Evaluation and application of RNA-Seq by MinION

    doi: 10.1093/dnares/dsy038

    Figure Lengend Snippet: Comparison of FL-cDNA-Seq and Illumina RNA-Seq. (A) The gene expression of FL-cDNA-Seq of LC2/ad (R9.4) was compared with that of TruSeq RNA (left) and SMART-Seq (right). Pearson correlation coefficients are shown on the graph. (B) Influence of sequencing depth on the estimation of gene expression level and gene detection. Reads for each method were randomly sampled in triplicate. The average of the Pearson correlation coefficients between TruSeq RNA and randomly sampled data for FL-cDNA-Seq and SMART-Seq is shown (left). The average number of genes with an expression level of more than 1 tpm or ppm is shown (right). (C) Comparison to qRT-qPCR. Forty-four genes detected by all methods were analyzed. The gene expression of these genes was normalized to GAPDH. Pearson correlation coefficients are shown on the graph. qRT-qPCR data of LC2/ad was obtained as in our previous study. 16

    Article Snippet: Optimization of the cDNA-Seq procedure Using the SMART-Seq v4 Ultra Low Input RNA Kit from Takara Bio, which is based on the Smart-Seq2 method, we synthesized full-length cDNA (FL-cDNA) from 50 ng total RNA isolated from seven lung adenocarcinoma-derived cell lines: PC-7, PC-9, H1975, H2228, VMRC-LCD, LC2/ad, and A549 ( ).

    Techniques: RNA Sequencing Assay, Expressing, Sequencing, Real-time Polymerase Chain Reaction

    Detection and determination of Has2 gene mutation A 454 bp region within exon2 of rat Has2 that contains the Cas9 mutation site was PCR amplified from genomic DNA of each cell clone as a template. The amplified product was gel-purified and then digested with Pas1. The final products were analyzed by agarose gel electrophoresis. DNA standards (std) are labeled. Panel A. Products amplified from RCS-o WT cells and clones 1 thru 7. Panel B. Pas1 digestion products of the amplified 454 bp products shown in panel A. Panel C. Products amplified from RCS-Cas9 WT cells and clones 3, 7, 39, 43, 53, 80 and 88. Panel D. Pas1 digestion products of the amplified 454 bp products shown in panel C. Panel E. Alignment of the targeted rat Has2 genomic sequences from RCS-Cas9 clone #7, allele 1 and allele 2 with the WT RCS-Cas9 cells. Deleted bases are indicated.

    Journal: Matrix biology : journal of the International Society for Matrix Biology

    Article Title: CRISPR/Cas9 knockout of HAS2 in rat chondrosarcoma chondrocytes demonstrates the requirement of hyaluronan for aggrecan retention

    doi: 10.1016/j.matbio.2016.04.002

    Figure Lengend Snippet: Detection and determination of Has2 gene mutation A 454 bp region within exon2 of rat Has2 that contains the Cas9 mutation site was PCR amplified from genomic DNA of each cell clone as a template. The amplified product was gel-purified and then digested with Pas1. The final products were analyzed by agarose gel electrophoresis. DNA standards (std) are labeled. Panel A. Products amplified from RCS-o WT cells and clones 1 thru 7. Panel B. Pas1 digestion products of the amplified 454 bp products shown in panel A. Panel C. Products amplified from RCS-Cas9 WT cells and clones 3, 7, 39, 43, 53, 80 and 88. Panel D. Pas1 digestion products of the amplified 454 bp products shown in panel C. Panel E. Alignment of the targeted rat Has2 genomic sequences from RCS-Cas9 clone #7, allele 1 and allele 2 with the WT RCS-Cas9 cells. Deleted bases are indicated.

    Article Snippet: To identify the mutation in the RCS-Cas9 clone #7, the same PCR product was also amplified but with a different DNA polymerase, TaKaRa LA Taq® DNA polymerase (Clontech), and this PCR product cloned into the pCR4-TOPO vector (Invitrogen) according to the manufacturer's instructions.

    Techniques: Mutagenesis, Polymerase Chain Reaction, Amplification, Purification, Agarose Gel Electrophoresis, Labeling, Clone Assay, Genomic Sequencing

    Screening of PAF-AH knockout transformants. (A) Physical map of homologous recombination. (B) Identification of transformants by PCR: 1, fragment of the hygromycin B resistance gene ( hyg ); 2, amplified fragment of PAF-AH from genomic DNA; 3, amplified T-DNA fragments adjusted to the right border of plasmid; 4, fragment amplified with primers PAF-AH-UP-F and Hyg-R. (C) Southern blot to verify T-DNA insertion copies of knockout transformants. The 450 bp fragment amplified from the hyg gene was labeled with 32 P-dCTP as probe 1. (D) Southern blot to confirm PAF-AH was replaced using the 500 bp fragment of PAF-AH as probe 2. WT, wild type strain; P, plasmid; KO15 and KO40: PAF-AH knockout transformants; M: DNA marker.

    Journal: PLoS ONE

    Article Title: Biological Role of Trichoderma harzianum-Derived Platelet-Activating Factor Acetylhydrolase (PAF-AH) on Stress Response and Antagonism

    doi: 10.1371/journal.pone.0100367

    Figure Lengend Snippet: Screening of PAF-AH knockout transformants. (A) Physical map of homologous recombination. (B) Identification of transformants by PCR: 1, fragment of the hygromycin B resistance gene ( hyg ); 2, amplified fragment of PAF-AH from genomic DNA; 3, amplified T-DNA fragments adjusted to the right border of plasmid; 4, fragment amplified with primers PAF-AH-UP-F and Hyg-R. (C) Southern blot to verify T-DNA insertion copies of knockout transformants. The 450 bp fragment amplified from the hyg gene was labeled with 32 P-dCTP as probe 1. (D) Southern blot to confirm PAF-AH was replaced using the 500 bp fragment of PAF-AH as probe 2. WT, wild type strain; P, plasmid; KO15 and KO40: PAF-AH knockout transformants; M: DNA marker.

    Article Snippet: Quantitative real-time PCR Total cDNA was synthesized from 1 µg wild type or other transformants' RNA using the PrimeScript RT Reagent kit with gDNA Eraser (Takara, Japan).

    Techniques: Knock-Out, Homologous Recombination, Polymerase Chain Reaction, Amplification, Plasmid Preparation, Southern Blot, Labeling, Marker

    Comparison of the BmP5CR1 genomic region between Daizo (+ Lg /+ Lg ) and DH6 ( Lg / Lg ). ( a ) Genomic PCR analysis of the three parts of BmP5CR1 by Ex or LA Taq polymerase HS. M is the DNA ladder marker. ( b ) Schematic representation of the BmP5CR1 gene structure. Lines and rectangular boxes show introns and exons, respectively. White boxes indicate the untranslated regions (UTRs), and grey boxes are the coding regions. Arrows indicate differences between Daizo and DH6. Arrowheads are primer sites used in this study. Numbers show the length (bp) of each region

    Journal: Heredity

    Article Title: Deficiency of a pyrroline-5-carboxylate reductase produces the yellowish green cocoon ‘Ryokuken’ of the silkworm, Bombyx mori

    doi: 10.1038/s41437-018-0051-8

    Figure Lengend Snippet: Comparison of the BmP5CR1 genomic region between Daizo (+ Lg /+ Lg ) and DH6 ( Lg / Lg ). ( a ) Genomic PCR analysis of the three parts of BmP5CR1 by Ex or LA Taq polymerase HS. M is the DNA ladder marker. ( b ) Schematic representation of the BmP5CR1 gene structure. Lines and rectangular boxes show introns and exons, respectively. White boxes indicate the untranslated regions (UTRs), and grey boxes are the coding regions. Arrows indicate differences between Daizo and DH6. Arrowheads are primer sites used in this study. Numbers show the length (bp) of each region

    Article Snippet: The region from transcription initiation to terminator (1F-9063R) and to the 1st intronic region on the Daizo genome (1F-5924R) of BmP5CR1 was amplified by 2.5 U Ex Taq polymerase or LA Taq polymerase (TaKaRa, Japan).

    Techniques: Polymerase Chain Reaction, Marker