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  • 94
    TaKaRa terra pcr direct polymerase mix
    Erh1 and Ccr4 regulate heterochromatin formation at rDNA repeats to protect rDNA integrity (A) ChIP-chip of Erh1-GFP (top) and myc-Ago1 (bottom) at rDNA repeats in WT. (B) ChIP-chip of H3K9me2 at rDNA repeats in WT and erh1 Δ (top), and in WT and ccr4 Δ (bottom). (C) ChIP-chip of H3K9me2 at rDNA repeats in WT and red1 Δ. (D) LEU2 and <t>ura4</t> + markers inserted at rDNA repeats (Ylp2.4pUC ura4 + -7) are de-repressed in erh1 Δ and ccr4 Δ. A schematic representation of Ylp2.4pUC ura4 + -7 is shown (top). WT and erh1 Δ (middle) or WT and ccr4 Δ (bottom) carrying Ylp2.4pUC ura4 + -7 were spotted onto complete medium or minimal medium lacking leucine and uracil (-Leu-Ura) and incubated at 30°C for 2 days. (E) Increased frequency of mitotic recombination at tandem rDNA repeats in erh1 Δ and ccr4 Δ. A schematic representation of rDNA :: ura4 + loss is shown (left). WT, erh1 Δ and ccr4 Δ carrying Ylp2.4pUC ura4 + -7 were first plated on medium lacking leucine, transferred to complete medium, and then replica plated to medium lacking uracil (-Ura). Mitotic recombination at rDNA loci was indicated by the lack of growth on -Ura plates, and was confirmed by genomic <t>PCR.</t> “ n .
    Terra Pcr Direct Polymerase Mix, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 172 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/terra pcr direct polymerase mix/product/TaKaRa
    Average 94 stars, based on 172 article reviews
    Price from $9.99 to $1999.99
    terra pcr direct polymerase mix - by Bioz Stars, 2020-05
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    90
    TaKaRa terra pcr direct genotyping kit
    Erh1 and Ccr4 regulate heterochromatin formation at rDNA repeats to protect rDNA integrity (A) ChIP-chip of Erh1-GFP (top) and myc-Ago1 (bottom) at rDNA repeats in WT. (B) ChIP-chip of H3K9me2 at rDNA repeats in WT and erh1 Δ (top), and in WT and ccr4 Δ (bottom). (C) ChIP-chip of H3K9me2 at rDNA repeats in WT and red1 Δ. (D) LEU2 and <t>ura4</t> + markers inserted at rDNA repeats (Ylp2.4pUC ura4 + -7) are de-repressed in erh1 Δ and ccr4 Δ. A schematic representation of Ylp2.4pUC ura4 + -7 is shown (top). WT and erh1 Δ (middle) or WT and ccr4 Δ (bottom) carrying Ylp2.4pUC ura4 + -7 were spotted onto complete medium or minimal medium lacking leucine and uracil (-Leu-Ura) and incubated at 30°C for 2 days. (E) Increased frequency of mitotic recombination at tandem rDNA repeats in erh1 Δ and ccr4 Δ. A schematic representation of rDNA :: ura4 + loss is shown (left). WT, erh1 Δ and ccr4 Δ carrying Ylp2.4pUC ura4 + -7 were first plated on medium lacking leucine, transferred to complete medium, and then replica plated to medium lacking uracil (-Ura). Mitotic recombination at rDNA loci was indicated by the lack of growth on -Ura plates, and was confirmed by genomic <t>PCR.</t> “ n .
    Terra Pcr Direct Genotyping Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/terra pcr direct genotyping kit/product/TaKaRa
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    terra pcr direct genotyping kit - by Bioz Stars, 2020-05
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    92
    TaKaRa terra pcr direct red dye premix
    Erh1 and Ccr4 regulate heterochromatin formation at rDNA repeats to protect rDNA integrity (A) ChIP-chip of Erh1-GFP (top) and myc-Ago1 (bottom) at rDNA repeats in WT. (B) ChIP-chip of H3K9me2 at rDNA repeats in WT and erh1 Δ (top), and in WT and ccr4 Δ (bottom). (C) ChIP-chip of H3K9me2 at rDNA repeats in WT and red1 Δ. (D) LEU2 and <t>ura4</t> + markers inserted at rDNA repeats (Ylp2.4pUC ura4 + -7) are de-repressed in erh1 Δ and ccr4 Δ. A schematic representation of Ylp2.4pUC ura4 + -7 is shown (top). WT and erh1 Δ (middle) or WT and ccr4 Δ (bottom) carrying Ylp2.4pUC ura4 + -7 were spotted onto complete medium or minimal medium lacking leucine and uracil (-Leu-Ura) and incubated at 30°C for 2 days. (E) Increased frequency of mitotic recombination at tandem rDNA repeats in erh1 Δ and ccr4 Δ. A schematic representation of rDNA :: ura4 + loss is shown (left). WT, erh1 Δ and ccr4 Δ carrying Ylp2.4pUC ura4 + -7 were first plated on medium lacking leucine, transferred to complete medium, and then replica plated to medium lacking uracil (-Ura). Mitotic recombination at rDNA loci was indicated by the lack of growth on -Ura plates, and was confirmed by genomic <t>PCR.</t> “ n .
    Terra Pcr Direct Red Dye Premix, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/terra pcr direct red dye premix/product/TaKaRa
    Average 92 stars, based on 19 article reviews
    Price from $9.99 to $1999.99
    terra pcr direct red dye premix - by Bioz Stars, 2020-05
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    94
    TaKaRa tb green qpcr mix
    Erh1 and Ccr4 regulate heterochromatin formation at rDNA repeats to protect rDNA integrity (A) ChIP-chip of Erh1-GFP (top) and myc-Ago1 (bottom) at rDNA repeats in WT. (B) ChIP-chip of H3K9me2 at rDNA repeats in WT and erh1 Δ (top), and in WT and ccr4 Δ (bottom). (C) ChIP-chip of H3K9me2 at rDNA repeats in WT and red1 Δ. (D) LEU2 and <t>ura4</t> + markers inserted at rDNA repeats (Ylp2.4pUC ura4 + -7) are de-repressed in erh1 Δ and ccr4 Δ. A schematic representation of Ylp2.4pUC ura4 + -7 is shown (top). WT and erh1 Δ (middle) or WT and ccr4 Δ (bottom) carrying Ylp2.4pUC ura4 + -7 were spotted onto complete medium or minimal medium lacking leucine and uracil (-Leu-Ura) and incubated at 30°C for 2 days. (E) Increased frequency of mitotic recombination at tandem rDNA repeats in erh1 Δ and ccr4 Δ. A schematic representation of rDNA :: ura4 + loss is shown (left). WT, erh1 Δ and ccr4 Δ carrying Ylp2.4pUC ura4 + -7 were first plated on medium lacking leucine, transferred to complete medium, and then replica plated to medium lacking uracil (-Ura). Mitotic recombination at rDNA loci was indicated by the lack of growth on -Ura plates, and was confirmed by genomic <t>PCR.</t> “ n .
    Tb Green Qpcr Mix, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Erh1 and Ccr4 regulate heterochromatin formation at rDNA repeats to protect rDNA integrity (A) ChIP-chip of Erh1-GFP (top) and myc-Ago1 (bottom) at rDNA repeats in WT. (B) ChIP-chip of H3K9me2 at rDNA repeats in WT and erh1 Δ (top), and in WT and ccr4 Δ (bottom). (C) ChIP-chip of H3K9me2 at rDNA repeats in WT and red1 Δ. (D) LEU2 and ura4 + markers inserted at rDNA repeats (Ylp2.4pUC ura4 + -7) are de-repressed in erh1 Δ and ccr4 Δ. A schematic representation of Ylp2.4pUC ura4 + -7 is shown (top). WT and erh1 Δ (middle) or WT and ccr4 Δ (bottom) carrying Ylp2.4pUC ura4 + -7 were spotted onto complete medium or minimal medium lacking leucine and uracil (-Leu-Ura) and incubated at 30°C for 2 days. (E) Increased frequency of mitotic recombination at tandem rDNA repeats in erh1 Δ and ccr4 Δ. A schematic representation of rDNA :: ura4 + loss is shown (left). WT, erh1 Δ and ccr4 Δ carrying Ylp2.4pUC ura4 + -7 were first plated on medium lacking leucine, transferred to complete medium, and then replica plated to medium lacking uracil (-Ura). Mitotic recombination at rDNA loci was indicated by the lack of growth on -Ura plates, and was confirmed by genomic PCR. “ n .

    Journal: Molecular cell

    Article Title: Enhancer of rudimentary cooperates with conserved RNA processing factors to promote meiotic mRNA decay and facultative heterochromatin assembly

    doi: 10.1016/j.molcel.2016.01.029

    Figure Lengend Snippet: Erh1 and Ccr4 regulate heterochromatin formation at rDNA repeats to protect rDNA integrity (A) ChIP-chip of Erh1-GFP (top) and myc-Ago1 (bottom) at rDNA repeats in WT. (B) ChIP-chip of H3K9me2 at rDNA repeats in WT and erh1 Δ (top), and in WT and ccr4 Δ (bottom). (C) ChIP-chip of H3K9me2 at rDNA repeats in WT and red1 Δ. (D) LEU2 and ura4 + markers inserted at rDNA repeats (Ylp2.4pUC ura4 + -7) are de-repressed in erh1 Δ and ccr4 Δ. A schematic representation of Ylp2.4pUC ura4 + -7 is shown (top). WT and erh1 Δ (middle) or WT and ccr4 Δ (bottom) carrying Ylp2.4pUC ura4 + -7 were spotted onto complete medium or minimal medium lacking leucine and uracil (-Leu-Ura) and incubated at 30°C for 2 days. (E) Increased frequency of mitotic recombination at tandem rDNA repeats in erh1 Δ and ccr4 Δ. A schematic representation of rDNA :: ura4 + loss is shown (left). WT, erh1 Δ and ccr4 Δ carrying Ylp2.4pUC ura4 + -7 were first plated on medium lacking leucine, transferred to complete medium, and then replica plated to medium lacking uracil (-Ura). Mitotic recombination at rDNA loci was indicated by the lack of growth on -Ura plates, and was confirmed by genomic PCR. “ n .

    Article Snippet: Loss of ura4 + was confirmed by PCR using Terra™ PCR Direct Polymerase Mix (TaKaRa Bio).

    Techniques: Chromatin Immunoprecipitation, Incubation, Polymerase Chain Reaction

    Schematic of the single-cell Quartz-Seq and Quartz-Chip methods . All of the steps of the whole-transcript amplification were executed in a single PCR tube. The first-strand cDNA was synthesized using the reverse transcription (RT) primer, which contains oligo-dT24, the T7 promoter (T7) and the PCR target region (M) sequences. After the first-strand synthesis, the majority of the RT primer was digested by exonuclease I, although it was not possible to eliminate the RT primer completely using this procedure. A poly-A tail was then added to the 3' ends of the first-strand cDNA and to any surviving RT primer. After the second-strand synthesis with the tagging primer, the resulting cDNA and the byproducts from the surviving primers contained the whole-transcript amplification (WTA) adaptor sequences, which include the RT primer sequence and the tagging primer sequence. These DNAs were used for the suppression PCR, which used the suppression PCR primer. Enrichment of the short DNA fragments, such as the byproducts, was suppressed. After the enrichment, the high-quality cDNA, which did not contain any byproducts, was obtained. The amplified cDNAs then had the T7 promoter sequence at the 3' ends of the DNA. These cDNAs were used for the Illumina sequencing and microarray experiments.

    Journal: Genome Biology

    Article Title: Quartz-Seq: a highly reproducible and sensitive single-cell RNA sequencing method, reveals non-genetic gene-expression heterogeneity

    doi: 10.1186/gb-2013-14-4-r31

    Figure Lengend Snippet: Schematic of the single-cell Quartz-Seq and Quartz-Chip methods . All of the steps of the whole-transcript amplification were executed in a single PCR tube. The first-strand cDNA was synthesized using the reverse transcription (RT) primer, which contains oligo-dT24, the T7 promoter (T7) and the PCR target region (M) sequences. After the first-strand synthesis, the majority of the RT primer was digested by exonuclease I, although it was not possible to eliminate the RT primer completely using this procedure. A poly-A tail was then added to the 3' ends of the first-strand cDNA and to any surviving RT primer. After the second-strand synthesis with the tagging primer, the resulting cDNA and the byproducts from the surviving primers contained the whole-transcript amplification (WTA) adaptor sequences, which include the RT primer sequence and the tagging primer sequence. These DNAs were used for the suppression PCR, which used the suppression PCR primer. Enrichment of the short DNA fragments, such as the byproducts, was suppressed. After the enrichment, the high-quality cDNA, which did not contain any byproducts, was obtained. The amplified cDNAs then had the T7 promoter sequence at the 3' ends of the DNA. These cDNAs were used for the Illumina sequencing and microarray experiments.

    Article Snippet: The use of this DNA polymerase (MightyAmp DNA Polymerase (Takara Bio, Inc., Tokyo, Japan), also marketed as Terra PCR Direct Polymerase (Clontech, Mountain View, CA, USA)) improved the yield of cDNA (see Additional file , Figure S2c and Figure S5) and the reproducibility of the WTA replication (see Additional file , Figure S2d and Figure S5).

    Techniques: Chromatin Immunoprecipitation, Amplification, Polymerase Chain Reaction, Synthesized, Sequencing, Microarray

    Klotho gene activation using a CRISPR NLuc knock-in HEK293 cell line. (A) Schematic view of the Klotho genomic area surrounding the 3′-UTR region, showing the stop codon, the sgRNA targets, and polyA tail. The P2A-NLuc sequence was inserted at the DSB site of the Klotho 3′-UTR.(B) PCR confirmation of positive clones using forward primer located at Klotho intron 4 (Int4) and NLuc reverse primer (NLrv) indicated in A. Cell line numbers are indicated. (C) Fold activation of Klotho gene expression by SAM evaluated with a CRISPR NLuc knock-in HEK293 line. Cells were analyzed by NLuc luciferase assay 2 days after transfection with dCas9-VP64, MS2-P65-HSF1 and the indicated sgRNA. Negative control: sgRNA cloning backbone empty vector. Positive control: Egr-1 transfected cells. Data are expressed as fold over negative control. Error bars show standard deviation among 6 replicates. Asterisks (*) indicate statistical significance of p

    Journal: Journal of molecular neuroscience : MN

    Article Title: Activation of the anti-aging and cognition-enhancing gene Klotho by CRISPR-dCas9 transcriptional effector complex

    doi: 10.1007/s12031-017-1011-0

    Figure Lengend Snippet: Klotho gene activation using a CRISPR NLuc knock-in HEK293 cell line. (A) Schematic view of the Klotho genomic area surrounding the 3′-UTR region, showing the stop codon, the sgRNA targets, and polyA tail. The P2A-NLuc sequence was inserted at the DSB site of the Klotho 3′-UTR.(B) PCR confirmation of positive clones using forward primer located at Klotho intron 4 (Int4) and NLuc reverse primer (NLrv) indicated in A. Cell line numbers are indicated. (C) Fold activation of Klotho gene expression by SAM evaluated with a CRISPR NLuc knock-in HEK293 line. Cells were analyzed by NLuc luciferase assay 2 days after transfection with dCas9-VP64, MS2-P65-HSF1 and the indicated sgRNA. Negative control: sgRNA cloning backbone empty vector. Positive control: Egr-1 transfected cells. Data are expressed as fold over negative control. Error bars show standard deviation among 6 replicates. Asterisks (*) indicate statistical significance of p

    Article Snippet: NLuc positive lines were confirmed by PCR using Terra PCR direct kit (Clontech) with the following primers: Klotho intron 4 forward: 5′-GTGTTGTGTGCAAAATACGTAATAA-3′; NLuc reverse: 5′-TGACATGGATGTCGATCTTCAG-3′.

    Techniques: Activation Assay, CRISPR, Knock-In, Sequencing, Polymerase Chain Reaction, Clone Assay, Expressing, Luciferase, Transfection, Negative Control, Plasmid Preparation, Positive Control, Standard Deviation

    Ectopic expression of OsWRKY42 suppresses salt and cellulase induced expression of JA biosynthesis and responsive genes in Arabidopsis transgenic lines. Expression of JA biosynthesis and response genes was measured in 35S::OsWRKY42 transgenic lines and wild type (Col-0) Arabidopsis plants after salt and cellulase treatment. a. For salt treatment, one-week old seedlings (wild type or 35S::OsWRKY42) were treated for 12 h with either MS or MS with 150 mM NaCl. b. For cellulase treatment, leaves of three weeks old Arabidopsis plants (wild type or 35S::OsWRKY42) were infiltrated with 2 U/ml of cellulase (Sigma) . Leaves were harvested after 12 h and processed for qPCR analysis. The graph represents the relative fold change using expression values from wild type or 35S::OsWRKY42 lines treated with water. c. Leaves of fourteen days old rice seedlings were syringe infiltrated with either LBA4404 or LBA4404/pH7FWG2::OsWRKY42. Twelve hours post treatment, leaves were harvested and processed for qRT-PCR. The relative fold change was calculated over control (leaves infiltrated only with LBA4404). AtUBQ5 and OsGAPDH were used as endogenous controls for rice and Arabidopsis, respectively. The average from three biological replicates is plotted on the graph. The error bar represents standard deviation. Data was analysed using the Student’s t -test for independent means. The asterisk above the bar indicates significant difference with p value

    Journal: BMC Plant Biology

    Article Title: Overexpression of a cell wall damage induced transcription factor, OsWRKY42, leads to enhanced callose deposition and tolerance to salt stress but does not enhance tolerance to bacterial infection

    doi: 10.1186/s12870-018-1391-5

    Figure Lengend Snippet: Ectopic expression of OsWRKY42 suppresses salt and cellulase induced expression of JA biosynthesis and responsive genes in Arabidopsis transgenic lines. Expression of JA biosynthesis and response genes was measured in 35S::OsWRKY42 transgenic lines and wild type (Col-0) Arabidopsis plants after salt and cellulase treatment. a. For salt treatment, one-week old seedlings (wild type or 35S::OsWRKY42) were treated for 12 h with either MS or MS with 150 mM NaCl. b. For cellulase treatment, leaves of three weeks old Arabidopsis plants (wild type or 35S::OsWRKY42) were infiltrated with 2 U/ml of cellulase (Sigma) . Leaves were harvested after 12 h and processed for qPCR analysis. The graph represents the relative fold change using expression values from wild type or 35S::OsWRKY42 lines treated with water. c. Leaves of fourteen days old rice seedlings were syringe infiltrated with either LBA4404 or LBA4404/pH7FWG2::OsWRKY42. Twelve hours post treatment, leaves were harvested and processed for qRT-PCR. The relative fold change was calculated over control (leaves infiltrated only with LBA4404). AtUBQ5 and OsGAPDH were used as endogenous controls for rice and Arabidopsis, respectively. The average from three biological replicates is plotted on the graph. The error bar represents standard deviation. Data was analysed using the Student’s t -test for independent means. The asterisk above the bar indicates significant difference with p value

    Article Snippet: The transgenic nature of the T1 plants was further confirmed by direct PCR (Terra PCR direct polymerase kit, Clontech using leaf tissue and sequencing of the amplified product.

    Techniques: Expressing, Transgenic Assay, Mass Spectrometry, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Standard Deviation

    Ectopic expression of OsWRKY42 enhances tolerance to salt stress in Arabidopsis transgenic lines. a. One-week-old TN-1 rice seedlings (n = 10) were dipped in each of the following: 150 mM NaCl solution, 20% ( w / v ) PEG-6000 and mock (water). Leaves were harvested 12 h post treatment and processed for qRT-PCR. Relative fold change was calculated over water treated samples and OsGAPDH was used as an internal control. b. For assaying salt tolerance, one-week-old Arabidopsis seedlings that are either constitutively expressing OsWRKY42 (35S::OsWRKY42) or wild type (Col-0) ( n = 20) were grown on MS agar medium with or without NaCl (150 mM, Merck). (c-d) The total fresh weight and number of root branches of individual seedlings was quantified on the fifteenth day post-treatment. e . For anthocyanin estimation, three weeks old Arabidopsis plants (wild type and 35S::OsWRKY42) were treated with either 150 mM NaCl or water. On the fifteenth day post treatment, leaves ( n = 3) were collected and anthocyanin was extracted in acidic methanol. Anthocyanin was estimated using a spectrophotometer and the anthocyanin content is expressed as absorbance per milligram of fresh weight (F.W). f . ROS estimation was done in detached leaves from adult plants. Leaf discs (n = 15) of either wild type or 35S::OsWRKY42 lines were placed on sterile filter papers soaked in MS solution either with or without NaCl (150 mM). 12 h post treatment, the leaves were stained with NBT. The O 2 − content was estimated spectrophotometrically and is expressed as change in absorbance per milligram of fresh weight. Leaves from three different plants were used as replicates. The data was analysed using one-way ANOVA and the Tukey-Karmer honestly significance difference test. The alphabets above the bar indicates significant difference with p value

    Journal: BMC Plant Biology

    Article Title: Overexpression of a cell wall damage induced transcription factor, OsWRKY42, leads to enhanced callose deposition and tolerance to salt stress but does not enhance tolerance to bacterial infection

    doi: 10.1186/s12870-018-1391-5

    Figure Lengend Snippet: Ectopic expression of OsWRKY42 enhances tolerance to salt stress in Arabidopsis transgenic lines. a. One-week-old TN-1 rice seedlings (n = 10) were dipped in each of the following: 150 mM NaCl solution, 20% ( w / v ) PEG-6000 and mock (water). Leaves were harvested 12 h post treatment and processed for qRT-PCR. Relative fold change was calculated over water treated samples and OsGAPDH was used as an internal control. b. For assaying salt tolerance, one-week-old Arabidopsis seedlings that are either constitutively expressing OsWRKY42 (35S::OsWRKY42) or wild type (Col-0) ( n = 20) were grown on MS agar medium with or without NaCl (150 mM, Merck). (c-d) The total fresh weight and number of root branches of individual seedlings was quantified on the fifteenth day post-treatment. e . For anthocyanin estimation, three weeks old Arabidopsis plants (wild type and 35S::OsWRKY42) were treated with either 150 mM NaCl or water. On the fifteenth day post treatment, leaves ( n = 3) were collected and anthocyanin was extracted in acidic methanol. Anthocyanin was estimated using a spectrophotometer and the anthocyanin content is expressed as absorbance per milligram of fresh weight (F.W). f . ROS estimation was done in detached leaves from adult plants. Leaf discs (n = 15) of either wild type or 35S::OsWRKY42 lines were placed on sterile filter papers soaked in MS solution either with or without NaCl (150 mM). 12 h post treatment, the leaves were stained with NBT. The O 2 − content was estimated spectrophotometrically and is expressed as change in absorbance per milligram of fresh weight. Leaves from three different plants were used as replicates. The data was analysed using one-way ANOVA and the Tukey-Karmer honestly significance difference test. The alphabets above the bar indicates significant difference with p value

    Article Snippet: The transgenic nature of the T1 plants was further confirmed by direct PCR (Terra PCR direct polymerase kit, Clontech using leaf tissue and sequencing of the amplified product.

    Techniques: Expressing, Transgenic Assay, Quantitative RT-PCR, Mass Spectrometry, Spectrophotometry, Staining