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ampl43  (Qiagen)


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    Structured Review

    Qiagen ampl43
    a) Schematic of <t>amPL43</t> expression vector for inducible CRISPRi library in S. cerevisiae . gRNA scaffold region contains a NotI site for cloning gRNA sequences, an RPR1 promoter with a TetO site, and a constant region for the gRNA. b) The expression fold-change of each target: ERG25 (three replicates), ERG11 (three replicates), and Sec14 (two replicates), as a result of gRNA induction by ATc, was calculated over time by qPCR. The mean for each sample is represented by a solid line. c) Schematic depicting the genomic region of PTA1 and ERV46. The gRNAs targeting the region between the two genes, depending on their proximity to each gene, could affect both genes. d) Histogram depicting the number of gRNAs per gene in two library replicates. The dashed blue line denotes the median: ~6. e) Scatter plot depicting the frequency of reads per gRNA between select biological replicates of the CRISPRi library. Pearson Correlation R value is reported for each pair.
    Ampl43, supplied by Qiagen, 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/product/ampl43/ampl43/bio_rxiv__2020__03__05__978619-142-4-7
    Average 90 stars, based on 1 article reviews
    ampl43 - by Bioz Stars, 2026-10
    90/100 stars

    Images

    1) Product Images from "An inducible CRISPR-interference library for genetic interrogation of Saccharomyces cerevisiae biology"

    Article Title: An inducible CRISPR-interference library for genetic interrogation of Saccharomyces cerevisiae biology

    Journal: bioRxiv

    doi: 10.1101/2020.03.05.978619

    a) Schematic of amPL43 expression vector for inducible CRISPRi library in S. cerevisiae . gRNA scaffold region contains a NotI site for cloning gRNA sequences, an RPR1 promoter with a TetO site, and a constant region for the gRNA. b) The expression fold-change of each target: ERG25 (three replicates), ERG11 (three replicates), and Sec14 (two replicates), as a result of gRNA induction by ATc, was calculated over time by qPCR. The mean for each sample is represented by a solid line. c) Schematic depicting the genomic region of PTA1 and ERV46. The gRNAs targeting the region between the two genes, depending on their proximity to each gene, could affect both genes. d) Histogram depicting the number of gRNAs per gene in two library replicates. The dashed blue line denotes the median: ~6. e) Scatter plot depicting the frequency of reads per gRNA between select biological replicates of the CRISPRi library. Pearson Correlation R value is reported for each pair.
    Figure Legend Snippet: a) Schematic of amPL43 expression vector for inducible CRISPRi library in S. cerevisiae . gRNA scaffold region contains a NotI site for cloning gRNA sequences, an RPR1 promoter with a TetO site, and a constant region for the gRNA. b) The expression fold-change of each target: ERG25 (three replicates), ERG11 (three replicates), and Sec14 (two replicates), as a result of gRNA induction by ATc, was calculated over time by qPCR. The mean for each sample is represented by a solid line. c) Schematic depicting the genomic region of PTA1 and ERV46. The gRNAs targeting the region between the two genes, depending on their proximity to each gene, could affect both genes. d) Histogram depicting the number of gRNAs per gene in two library replicates. The dashed blue line denotes the median: ~6. e) Scatter plot depicting the frequency of reads per gRNA between select biological replicates of the CRISPRi library. Pearson Correlation R value is reported for each pair.

    Techniques Used: Expressing, Plasmid Preparation, Clone Assay

    Related Articles

    Expressing:

    Article Title: An inducible CRISPR-interference library for genetic interrogation of Saccharomyces cerevisiae biology
    Article Snippet: library spacers were amplified and extended as described by Smith et al. ( ). .. To generate the library, amPL43 was maxiprepped (Qiagen, Hilden, Germany) and cut with NotI. .. Gibson Assembly was used to clone the oligos in the NotI site of amPL43.

    Plasmid Preparation:

    Article Title: An inducible CRISPR-interference library for genetic interrogation of Saccharomyces cerevisiae biology
    Article Snippet: library spacers were amplified and extended as described by Smith et al. ( ). .. To generate the library, amPL43 was maxiprepped (Qiagen, Hilden, Germany) and cut with NotI. .. Gibson Assembly was used to clone the oligos in the NotI site of amPL43.

    Clone Assay:

    Article Title: An inducible CRISPR-interference library for genetic interrogation of Saccharomyces cerevisiae biology
    Article Snippet: library spacers were amplified and extended as described by Smith et al. ( ). .. To generate the library, amPL43 was maxiprepped (Qiagen, Hilden, Germany) and cut with NotI. .. Gibson Assembly was used to clone the oligos in the NotI site of amPL43.

    Synthesized:

    Article Title: An inducible CRISPR-interference library for genetic interrogation of Saccharomyces cerevisiae biology
    Article Snippet: library spacers were amplified and extended as described by Smith et al. ( ). .. To generate the library, amPL43 was maxiprepped (Qiagen, Hilden, Germany) and cut with NotI. .. Gibson Assembly was used to clone the oligos in the NotI site of amPL43.



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    a Schematic of <t>amPL43</t> expression vector for inducible CRISPRi library in S. cerevisiae . b The expression fold change of each target: ERG25 (three replicates), ERG11 (five replicates), and Sec14 (four replicates), as a result of presence of sgRNA, without induction after 24 hours, as measured by qPCR. The mean for each sample is represented by a solid line. c The expression fold change of each target: ERG25 (three replicates), ERG11 (three replicates), and Sec14 (two replicates), as a result of gRNA induction by ATc, was calculated over time by qPCR. The mean for each sample is represented by a solid line. d Schematic depicting the genomic region of PTA1 and ERV46. The gRNAs targeting the region between the two genes, depending on their proximity to each gene, could affect both genes. e Histogram depicting the number of gRNAs per gene in two library replicates. The dashed blue line denotes the median: ~6. f Scatter plot depicting the frequency of reads per gRNA between select biological replicates of the CRISPRi library. Pearson Correlation R value is reported for each pair.
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    a) Schematic of <t>amPL43</t> expression vector for inducible CRISPRi library in S. cerevisiae . gRNA scaffold region contains a NotI site for cloning gRNA sequences, an RPR1 promoter with a TetO site, and a constant region for the gRNA. b) The expression fold-change of each target: ERG25 (three replicates), ERG11 (three replicates), and Sec14 (two replicates), as a result of gRNA induction by ATc, was calculated over time by qPCR. The mean for each sample is represented by a solid line. c) Schematic depicting the genomic region of PTA1 and ERV46. The gRNAs targeting the region between the two genes, depending on their proximity to each gene, could affect both genes. d) Histogram depicting the number of gRNAs per gene in two library replicates. The dashed blue line denotes the median: ~6. e) Scatter plot depicting the frequency of reads per gRNA between select biological replicates of the CRISPRi library. Pearson Correlation R value is reported for each pair.
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    Image Search Results


    a Schematic of amPL43 expression vector for inducible CRISPRi library in S. cerevisiae . b The expression fold change of each target: ERG25 (three replicates), ERG11 (five replicates), and Sec14 (four replicates), as a result of presence of sgRNA, without induction after 24 hours, as measured by qPCR. The mean for each sample is represented by a solid line. c The expression fold change of each target: ERG25 (three replicates), ERG11 (three replicates), and Sec14 (two replicates), as a result of gRNA induction by ATc, was calculated over time by qPCR. The mean for each sample is represented by a solid line. d Schematic depicting the genomic region of PTA1 and ERV46. The gRNAs targeting the region between the two genes, depending on their proximity to each gene, could affect both genes. e Histogram depicting the number of gRNAs per gene in two library replicates. The dashed blue line denotes the median: ~6. f Scatter plot depicting the frequency of reads per gRNA between select biological replicates of the CRISPRi library. Pearson Correlation R value is reported for each pair.

    Journal: Communications Biology

    Article Title: An inducible CRISPR interference library for genetic interrogation of Saccharomyces cerevisiae biology

    doi: 10.1038/s42003-020-01452-9

    Figure Lengend Snippet: a Schematic of amPL43 expression vector for inducible CRISPRi library in S. cerevisiae . b The expression fold change of each target: ERG25 (three replicates), ERG11 (five replicates), and Sec14 (four replicates), as a result of presence of sgRNA, without induction after 24 hours, as measured by qPCR. The mean for each sample is represented by a solid line. c The expression fold change of each target: ERG25 (three replicates), ERG11 (three replicates), and Sec14 (two replicates), as a result of gRNA induction by ATc, was calculated over time by qPCR. The mean for each sample is represented by a solid line. d Schematic depicting the genomic region of PTA1 and ERV46. The gRNAs targeting the region between the two genes, depending on their proximity to each gene, could affect both genes. e Histogram depicting the number of gRNAs per gene in two library replicates. The dashed blue line denotes the median: ~6. f Scatter plot depicting the frequency of reads per gRNA between select biological replicates of the CRISPRi library. Pearson Correlation R value is reported for each pair.

    Article Snippet: AmPl43 was also deposited to Addgene (Addgene ID: 161830).

    Techniques: Expressing, Plasmid Preparation

    a) Schematic of amPL43 expression vector for inducible CRISPRi library in S. cerevisiae . gRNA scaffold region contains a NotI site for cloning gRNA sequences, an RPR1 promoter with a TetO site, and a constant region for the gRNA. b) The expression fold-change of each target: ERG25 (three replicates), ERG11 (three replicates), and Sec14 (two replicates), as a result of gRNA induction by ATc, was calculated over time by qPCR. The mean for each sample is represented by a solid line. c) Schematic depicting the genomic region of PTA1 and ERV46. The gRNAs targeting the region between the two genes, depending on their proximity to each gene, could affect both genes. d) Histogram depicting the number of gRNAs per gene in two library replicates. The dashed blue line denotes the median: ~6. e) Scatter plot depicting the frequency of reads per gRNA between select biological replicates of the CRISPRi library. Pearson Correlation R value is reported for each pair.

    Journal: bioRxiv

    Article Title: An inducible CRISPR-interference library for genetic interrogation of Saccharomyces cerevisiae biology

    doi: 10.1101/2020.03.05.978619

    Figure Lengend Snippet: a) Schematic of amPL43 expression vector for inducible CRISPRi library in S. cerevisiae . gRNA scaffold region contains a NotI site for cloning gRNA sequences, an RPR1 promoter with a TetO site, and a constant region for the gRNA. b) The expression fold-change of each target: ERG25 (three replicates), ERG11 (three replicates), and Sec14 (two replicates), as a result of gRNA induction by ATc, was calculated over time by qPCR. The mean for each sample is represented by a solid line. c) Schematic depicting the genomic region of PTA1 and ERV46. The gRNAs targeting the region between the two genes, depending on their proximity to each gene, could affect both genes. d) Histogram depicting the number of gRNAs per gene in two library replicates. The dashed blue line denotes the median: ~6. e) Scatter plot depicting the frequency of reads per gRNA between select biological replicates of the CRISPRi library. Pearson Correlation R value is reported for each pair.

    Article Snippet: To generate the library, amPL43 was maxiprepped (Qiagen, Hilden, Germany) and cut with NotI.

    Techniques: Expressing, Plasmid Preparation, Clone Assay