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undigested pcr product  (Qiagen)


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    Qiagen undigested pcr product
    Undigested Pcr Product, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 61684 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/undigested+pcr+product/QIAquick+PCR+Purification+Kit/pm30778069-328-21-28
    Average 99 stars, based on 61684 article reviews
    undigested pcr product - by Bioz Stars, 2026-10
    99/100 stars

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    Related Articles

    CRISPR:

    Article Title: Precise tuning of gene expression levels in mammalian cells.
    Article Snippet: The PCRs were purified using the MinElute PCR Purification Kit (Qiagen). .. To enrich for CRISPR/Cas9-edited gDNA/cDNA, we digested WT DNA from these PCR products with SacI-HF (NEB) and gelpurified the remaining CRISPR/Cas9-modified, undigested PCR product (QIAquick Gel Extraction Kit, Qiagen). .. Illumina sequencing adaptors and unique barcode combinations were appended to the HDR-enriched DNA by PCR.

    Article Title: Precise tuning of gene expression levels in mammalian cells
    Article Snippet: The PCRs were purified using the MinElute PCR Purification Kit (Qiagen). .. To enrich for CRISPR/Cas9-edited gDNA/cDNA, we digested WT DNA from these PCR products with SacI-HF (NEB) and gel-purified the remaining CRISPR/Cas9-modified, undigested PCR product (QIAquick Gel Extraction Kit, Qiagen). .. Illumina sequencing adaptors and unique barcode combinations were appended to the HDR-enriched DNA by PCR.

    Polymerase Chain Reaction:

    Article Title: Precise tuning of gene expression levels in mammalian cells.
    Article Snippet: The PCRs were purified using the MinElute PCR Purification Kit (Qiagen). .. To enrich for CRISPR/Cas9-edited gDNA/cDNA, we digested WT DNA from these PCR products with SacI-HF (NEB) and gelpurified the remaining CRISPR/Cas9-modified, undigested PCR product (QIAquick Gel Extraction Kit, Qiagen). .. Illumina sequencing adaptors and unique barcode combinations were appended to the HDR-enriched DNA by PCR.

    Article Title: Precise tuning of gene expression levels in mammalian cells
    Article Snippet: The PCRs were purified using the MinElute PCR Purification Kit (Qiagen). .. To enrich for CRISPR/Cas9-edited gDNA/cDNA, we digested WT DNA from these PCR products with SacI-HF (NEB) and gel-purified the remaining CRISPR/Cas9-modified, undigested PCR product (QIAquick Gel Extraction Kit, Qiagen). .. Illumina sequencing adaptors and unique barcode combinations were appended to the HDR-enriched DNA by PCR.

    Gel Extraction:

    Article Title: Precise tuning of gene expression levels in mammalian cells.
    Article Snippet: The PCRs were purified using the MinElute PCR Purification Kit (Qiagen). .. To enrich for CRISPR/Cas9-edited gDNA/cDNA, we digested WT DNA from these PCR products with SacI-HF (NEB) and gelpurified the remaining CRISPR/Cas9-modified, undigested PCR product (QIAquick Gel Extraction Kit, Qiagen). .. Illumina sequencing adaptors and unique barcode combinations were appended to the HDR-enriched DNA by PCR.

    Article Title: Precise tuning of gene expression levels in mammalian cells
    Article Snippet: The PCRs were purified using the MinElute PCR Purification Kit (Qiagen). .. To enrich for CRISPR/Cas9-edited gDNA/cDNA, we digested WT DNA from these PCR products with SacI-HF (NEB) and gel-purified the remaining CRISPR/Cas9-modified, undigested PCR product (QIAquick Gel Extraction Kit, Qiagen). .. Illumina sequencing adaptors and unique barcode combinations were appended to the HDR-enriched DNA by PCR.



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    Combination of ZFN mRNA and AAV6 vectors promotes high levels of gene correction-like events at the CCR5 locus in CD8 + T cells. ( A ) Schematic showing use of AAV vector as a template for homology directed repair (HDR) of a double-strand break (DSB), as induced by target-specific nucleases, to introduce a corrected gene sequence or transgene into a predetermined genomic target site. ( B ) Schematic of AAV vector genomes containing CCR5 -RFLP homology donors. L (left) and R (right) refer to CCR5 genomic sequences, comprising 509 and 1349 bp respectively. ( C ) CD8 + T cells were transduced with AAV6 vectors carrying the CCR5 -RFLP donor at indicated doses for 16 hours, then electroporated with CCR5 ZFN mRNA (60μg/ml). Cells were analyzed 7 days post-electroporation by deep <t>sequencing</t> to measure the efficiency of genome modification (% indels and RFLP). Combined results of three experiments using three different CD8 + T cell donors are shown. Mean ± SD. * P < 0.05, two-tailed t -test to compare %RFLP between conditions with different doses of AAV6 donor in the presence of ZFN mRNA treatment. ( D ) Dose-dependent insertion of XhoI site at CCR5 , confirmed by RFLP analysis. Adjusted%RFLP HDR are shown below the lanes with visible RFLP bands based on deep sequencing results of the undigested top band due to incomplete digestion. The %RFLP before adjustment are shown in parentheses. One representative experiment is shown.
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    Image Search Results


    Combination of ZFN mRNA and AAV6 vectors promotes high levels of gene correction-like events at the CCR5 locus in CD8 + T cells. ( A ) Schematic showing use of AAV vector as a template for homology directed repair (HDR) of a double-strand break (DSB), as induced by target-specific nucleases, to introduce a corrected gene sequence or transgene into a predetermined genomic target site. ( B ) Schematic of AAV vector genomes containing CCR5 -RFLP homology donors. L (left) and R (right) refer to CCR5 genomic sequences, comprising 509 and 1349 bp respectively. ( C ) CD8 + T cells were transduced with AAV6 vectors carrying the CCR5 -RFLP donor at indicated doses for 16 hours, then electroporated with CCR5 ZFN mRNA (60μg/ml). Cells were analyzed 7 days post-electroporation by deep sequencing to measure the efficiency of genome modification (% indels and RFLP). Combined results of three experiments using three different CD8 + T cell donors are shown. Mean ± SD. * P < 0.05, two-tailed t -test to compare %RFLP between conditions with different doses of AAV6 donor in the presence of ZFN mRNA treatment. ( D ) Dose-dependent insertion of XhoI site at CCR5 , confirmed by RFLP analysis. Adjusted%RFLP HDR are shown below the lanes with visible RFLP bands based on deep sequencing results of the undigested top band due to incomplete digestion. The %RFLP before adjustment are shown in parentheses. One representative experiment is shown.

    Journal: Nucleic Acids Research

    Article Title: Highly efficient homology-driven genome editing in human T cells by combining zinc-finger nuclease mRNA and AAV6 donor delivery

    doi: 10.1093/nar/gkv1121

    Figure Lengend Snippet: Combination of ZFN mRNA and AAV6 vectors promotes high levels of gene correction-like events at the CCR5 locus in CD8 + T cells. ( A ) Schematic showing use of AAV vector as a template for homology directed repair (HDR) of a double-strand break (DSB), as induced by target-specific nucleases, to introduce a corrected gene sequence or transgene into a predetermined genomic target site. ( B ) Schematic of AAV vector genomes containing CCR5 -RFLP homology donors. L (left) and R (right) refer to CCR5 genomic sequences, comprising 509 and 1349 bp respectively. ( C ) CD8 + T cells were transduced with AAV6 vectors carrying the CCR5 -RFLP donor at indicated doses for 16 hours, then electroporated with CCR5 ZFN mRNA (60μg/ml). Cells were analyzed 7 days post-electroporation by deep sequencing to measure the efficiency of genome modification (% indels and RFLP). Combined results of three experiments using three different CD8 + T cell donors are shown. Mean ± SD. * P < 0.05, two-tailed t -test to compare %RFLP between conditions with different doses of AAV6 donor in the presence of ZFN mRNA treatment. ( D ) Dose-dependent insertion of XhoI site at CCR5 , confirmed by RFLP analysis. Adjusted%RFLP HDR are shown below the lanes with visible RFLP bands based on deep sequencing results of the undigested top band due to incomplete digestion. The %RFLP before adjustment are shown in parentheses. One representative experiment is shown.

    Article Snippet: For Illumina deep sequencing, gel-purified Out-Out1 PCR products (undigested band or untreated) were amplified with a target-specific Miseq adaptor primer pair (Supplementary Table S1, adaptor primers) and sequence barcodes were added in the subsequent PCR reaction using the barcode primer pairs.

    Techniques: Plasmid Preparation, Introduce, Sequencing, Genomic Sequencing, Transduction, Electroporation, Modification, Two Tailed Test

    Combination of ZFN mRNA and AAV6 vectors promotes high levels of transgene addition at the CCR5 locus in CD8 + T cells. ( A ) Schematic of AAV vector genomes containing CCR5 -GFP homology donors. L (left) and R (right) refer to CCR5 genomic sequences, comprising 476 and 1428 bp, respectively. ( B ) CD8 + T cells were treated as described in Figure , but using CCR5 -GFP donor vectors, with and without CCR5 ZFN mRNA electroporation. Cells were collected at 14 days post-transduction and analyzed by flow cytometry for %GFP + , and by deep sequencing to measure% indels. Results were combined from three experiments using three different CD8+ T cell donors. Mean ± SD. * P < 0.05, two-tailed t -test to compare%GFP + between conditions with different doses of AAV6 donor in the presence of ZFN mRNA treatment. ( C ) Flow cytometry plots from one representative experiment were shown using 1 × 10 6 vg/cell CCR5 -GFP donor, at 14 days post-electroporation. ( D ) Confirmation of targeted integration of GFP expression cassette at the CCR5 locus by semi-quantitative PCR. The %GFP HDR was estimated by comparison to standards.

    Journal: Nucleic Acids Research

    Article Title: Highly efficient homology-driven genome editing in human T cells by combining zinc-finger nuclease mRNA and AAV6 donor delivery

    doi: 10.1093/nar/gkv1121

    Figure Lengend Snippet: Combination of ZFN mRNA and AAV6 vectors promotes high levels of transgene addition at the CCR5 locus in CD8 + T cells. ( A ) Schematic of AAV vector genomes containing CCR5 -GFP homology donors. L (left) and R (right) refer to CCR5 genomic sequences, comprising 476 and 1428 bp, respectively. ( B ) CD8 + T cells were treated as described in Figure , but using CCR5 -GFP donor vectors, with and without CCR5 ZFN mRNA electroporation. Cells were collected at 14 days post-transduction and analyzed by flow cytometry for %GFP + , and by deep sequencing to measure% indels. Results were combined from three experiments using three different CD8+ T cell donors. Mean ± SD. * P < 0.05, two-tailed t -test to compare%GFP + between conditions with different doses of AAV6 donor in the presence of ZFN mRNA treatment. ( C ) Flow cytometry plots from one representative experiment were shown using 1 × 10 6 vg/cell CCR5 -GFP donor, at 14 days post-electroporation. ( D ) Confirmation of targeted integration of GFP expression cassette at the CCR5 locus by semi-quantitative PCR. The %GFP HDR was estimated by comparison to standards.

    Article Snippet: For Illumina deep sequencing, gel-purified Out-Out1 PCR products (undigested band or untreated) were amplified with a target-specific Miseq adaptor primer pair (Supplementary Table S1, adaptor primers) and sequence barcodes were added in the subsequent PCR reaction using the barcode primer pairs.

    Techniques: Plasmid Preparation, Genomic Sequencing, Electroporation, Transduction, Flow Cytometry, Sequencing, Two Tailed Test, Expressing, Real-time Polymerase Chain Reaction, Comparison

    AAV6 vectors and ZFN mRNA promote high levels of gene correction-like events at the AAVS1 locus in CD8 + T cells. ( A ) Schematics of AAV6 vectors used to deliver AAVS1 homology donors. L (left) and R (right) refer to AAVS1 genomic sequences, comprising 789 and 837 bp respectively . ( B ) CD8 + T cells were transduced with AAV6 vectors carrying the AAVS1 -RFLP donor at indicated doses (vg/cell) for 16 h and/or electroporated with AAVS1 ZFN mRNA. Cells were analyzed 7 days post-electroporation by Illumina deep sequencing to measure the efficiency of genome modification (% indels and RFLP). Results are combined from three experiments using three different CD8 + T cell donors. Mean ± SD. * P < 0.05, two-tailed t -test to compare%RFLP between conditions with different doses of AAV6 donor in the presence of ZFN mRNA treatment. ( C ) Confirmation of insertion of HindIII site at the AAVS1 locus by RFLP assay. Adjusted%RFLP HDR are shown below the lanes with visible RFLP bands based on deep sequencing results of the undigested top band due to incomplete digestion. The %RFLP before adjustment are shown in parentheses. One representative experiment is shown.

    Journal: Nucleic Acids Research

    Article Title: Highly efficient homology-driven genome editing in human T cells by combining zinc-finger nuclease mRNA and AAV6 donor delivery

    doi: 10.1093/nar/gkv1121

    Figure Lengend Snippet: AAV6 vectors and ZFN mRNA promote high levels of gene correction-like events at the AAVS1 locus in CD8 + T cells. ( A ) Schematics of AAV6 vectors used to deliver AAVS1 homology donors. L (left) and R (right) refer to AAVS1 genomic sequences, comprising 789 and 837 bp respectively . ( B ) CD8 + T cells were transduced with AAV6 vectors carrying the AAVS1 -RFLP donor at indicated doses (vg/cell) for 16 h and/or electroporated with AAVS1 ZFN mRNA. Cells were analyzed 7 days post-electroporation by Illumina deep sequencing to measure the efficiency of genome modification (% indels and RFLP). Results are combined from three experiments using three different CD8 + T cell donors. Mean ± SD. * P < 0.05, two-tailed t -test to compare%RFLP between conditions with different doses of AAV6 donor in the presence of ZFN mRNA treatment. ( C ) Confirmation of insertion of HindIII site at the AAVS1 locus by RFLP assay. Adjusted%RFLP HDR are shown below the lanes with visible RFLP bands based on deep sequencing results of the undigested top band due to incomplete digestion. The %RFLP before adjustment are shown in parentheses. One representative experiment is shown.

    Article Snippet: For Illumina deep sequencing, gel-purified Out-Out1 PCR products (undigested band or untreated) were amplified with a target-specific Miseq adaptor primer pair (Supplementary Table S1, adaptor primers) and sequence barcodes were added in the subsequent PCR reaction using the barcode primer pairs.

    Techniques: Genomic Sequencing, Transduction, Electroporation, Sequencing, Modification, Two Tailed Test, RFLP Assay

    AAV6 vectors and ZFN mRNA promote high levels of transgene addition at the AAVS1 locus in CD8 + T cells. ( A ) Schematics of AAV6 vectors used to deliver AAVS1 -GFP homology donors. L (left) and R (right) refer to AAVS1 genomic sequences, comprising 789 and 570 bp, respectively. ( B ) CD8 + T cells were treated as described in Figure , but using AAVS1 -GFP donor. Cells were collected 14 days post-transduction and analyzed by flow cytometry for %GFP + , and by deep sequencing to measure% indels. Results were combined from three experiments using three different CD8 + T cell donors. Mean ± SD. * P < 0.05, two-tailed t -test to compare %GFP + between conditions with different doses of AAV6 donor in the presence of ZFN mRNA treatment. ( C ) Flow cytometry plots from a representative experiment using 1 × 10 6 vg/cell AAVS1 -GFP donor at 14 days post-electroporation.

    Journal: Nucleic Acids Research

    Article Title: Highly efficient homology-driven genome editing in human T cells by combining zinc-finger nuclease mRNA and AAV6 donor delivery

    doi: 10.1093/nar/gkv1121

    Figure Lengend Snippet: AAV6 vectors and ZFN mRNA promote high levels of transgene addition at the AAVS1 locus in CD8 + T cells. ( A ) Schematics of AAV6 vectors used to deliver AAVS1 -GFP homology donors. L (left) and R (right) refer to AAVS1 genomic sequences, comprising 789 and 570 bp, respectively. ( B ) CD8 + T cells were treated as described in Figure , but using AAVS1 -GFP donor. Cells were collected 14 days post-transduction and analyzed by flow cytometry for %GFP + , and by deep sequencing to measure% indels. Results were combined from three experiments using three different CD8 + T cell donors. Mean ± SD. * P < 0.05, two-tailed t -test to compare %GFP + between conditions with different doses of AAV6 donor in the presence of ZFN mRNA treatment. ( C ) Flow cytometry plots from a representative experiment using 1 × 10 6 vg/cell AAVS1 -GFP donor at 14 days post-electroporation.

    Article Snippet: For Illumina deep sequencing, gel-purified Out-Out1 PCR products (undigested band or untreated) were amplified with a target-specific Miseq adaptor primer pair (Supplementary Table S1, adaptor primers) and sequence barcodes were added in the subsequent PCR reaction using the barcode primer pairs.

    Techniques: Genomic Sequencing, Transduction, Flow Cytometry, Sequencing, Two Tailed Test, Electroporation