engen sgrna synthesis kit  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    New England Biolabs engen sgrna synthesis kit
    Engen Sgrna Synthesis Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen sgrna synthesis kit/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen sgrna synthesis kit - by Bioz Stars, 2023-02
    86/100 stars

    Images

    engen sgrna synthesis kit  (New England Biolabs)


    Bioz Verified Symbol New England Biolabs is a verified supplier
    Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    New England Biolabs engen sgrna synthesis kit
    Engen Sgrna Synthesis Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen sgrna synthesis kit/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen sgrna synthesis kit - by Bioz Stars, 2023-02
    86/100 stars

    Images

    engen lba cas12a  (New England Biolabs)


    Bioz Verified Symbol New England Biolabs is a verified supplier
    Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    New England Biolabs engen lba cas12a
    Engen Lba Cas12a, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen lba cas12a/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen lba cas12a - by Bioz Stars, 2023-02
    86/100 stars

    Images

    engen lba cas12a cpf1 kit  (New England Biolabs)


    Bioz Verified Symbol New England Biolabs is a verified supplier
    Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    New England Biolabs engen lba cas12a cpf1 kit
    Feasibility of <t>Cas12a–PMNT-based</t> visual method for nucleic acid detection (1) NEBuffer + positive plasmid DNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (2) NEBuffer + positive plasmid DNA + crRNA + Rnase Inhibitor + ssDNA + PMNT; (3) NEBuffer + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (4) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + PMNT; (5) NEBuffer + ssDNA + PMNT; (6) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (7) NEBuffer + negative plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (8) S1 Reaction Buffer + S1 Nuclease + ssDNA + PMNT.
    Engen Lba Cas12a Cpf1 Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen lba cas12a cpf1 kit/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen lba cas12a cpf1 kit - by Bioz Stars, 2023-02
    86/100 stars

    Images

    1) Product Images from "A Rapid and Visual Method for Nucleic Acid Detection of Escherichia coli O157:H7 Based on CRISPR/Cas12a-PMNT"

    Article Title: A Rapid and Visual Method for Nucleic Acid Detection of Escherichia coli O157:H7 Based on CRISPR/Cas12a-PMNT

    Journal: Foods

    doi: 10.3390/foods12020236

    Feasibility of Cas12a–PMNT-based visual method for nucleic acid detection (1) NEBuffer + positive plasmid DNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (2) NEBuffer + positive plasmid DNA + crRNA + Rnase Inhibitor + ssDNA + PMNT; (3) NEBuffer + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (4) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + PMNT; (5) NEBuffer + ssDNA + PMNT; (6) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (7) NEBuffer + negative plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (8) S1 Reaction Buffer + S1 Nuclease + ssDNA + PMNT.
    Figure Legend Snippet: Feasibility of Cas12a–PMNT-based visual method for nucleic acid detection (1) NEBuffer + positive plasmid DNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (2) NEBuffer + positive plasmid DNA + crRNA + Rnase Inhibitor + ssDNA + PMNT; (3) NEBuffer + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (4) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + PMNT; (5) NEBuffer + ssDNA + PMNT; (6) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (7) NEBuffer + negative plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (8) S1 Reaction Buffer + S1 Nuclease + ssDNA + PMNT.

    Techniques Used: Plasmid Preparation

    engen sgrna synthesis kit  (New England Biolabs)


    Bioz Verified Symbol New England Biolabs is a verified supplier
    Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    New England Biolabs engen sgrna synthesis kit
    Engen Sgrna Synthesis Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen sgrna synthesis kit/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen sgrna synthesis kit - by Bioz Stars, 2023-02
    86/100 stars

    Images

    engen sgrna synthesis kit  (New England Biolabs)


    Bioz Verified Symbol New England Biolabs is a verified supplier
    Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    New England Biolabs engen sgrna synthesis kit
    Engen Sgrna Synthesis Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen sgrna synthesis kit/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen sgrna synthesis kit - by Bioz Stars, 2023-02
    86/100 stars

    Images

    engen sgrna enzyme mix  (New England Biolabs)


    Bioz Verified Symbol New England Biolabs is a verified supplier
    Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    New England Biolabs engen sgrna enzyme mix
    Engen Sgrna Enzyme Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen sgrna enzyme mix/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen sgrna enzyme mix - by Bioz Stars, 2023-02
    86/100 stars

    Images

    engen 2× sgrna reaction mix  (New England Biolabs)


    Bioz Verified Symbol New England Biolabs is a verified supplier
    Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    New England Biolabs engen 2× sgrna reaction mix
    Single guide RNA sequences and primers used for amplification and sequencing of target genomic regions.
    Engen 2× Sgrna Reaction Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen 2× sgrna reaction mix/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen 2× sgrna reaction mix - by Bioz Stars, 2023-02
    86/100 stars

    Images

    1) Product Images from "CRISPR/Cas9-Mediated Gene Editing in Salmonids Cells and Efficient Establishment of Edited Clonal Cell Lines"

    Article Title: CRISPR/Cas9-Mediated Gene Editing in Salmonids Cells and Efficient Establishment of Edited Clonal Cell Lines

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms232416218

    Single guide RNA sequences and primers used for amplification and sequencing of target genomic regions.
    Figure Legend Snippet: Single guide RNA sequences and primers used for amplification and sequencing of target genomic regions.

    Techniques Used: Amplification, Sequencing

    sgRNA analysis and in vitro cleavage assay for cr2 and mmp9 genes. ( a ) sgRNA analysis on 2% agarose gel. UD: undenatured sgRNA, D: denatured sgRNA. Single bands detected for the denatured sgRNAs highlighted the purity of the products, and multiple bands in the undenatured sgRNA resulted from the presence of RNA secondary structures; ( b ) gel shows duplicate lanes for each sample. Lanes 1–6: experimental samples, lane 7: no Cas9 control, lane 8: no sgRNA control; ( c ) lane 1: no Cas9 control, lane 2: no sgRNA control, lanes 3–5: experimental samples. Cleavage products at expected band sizes were detected for all samples containing the sgRNA + Cas9 + gDNA. No product at the expected band size was observed for all the control samples.
    Figure Legend Snippet: sgRNA analysis and in vitro cleavage assay for cr2 and mmp9 genes. ( a ) sgRNA analysis on 2% agarose gel. UD: undenatured sgRNA, D: denatured sgRNA. Single bands detected for the denatured sgRNAs highlighted the purity of the products, and multiple bands in the undenatured sgRNA resulted from the presence of RNA secondary structures; ( b ) gel shows duplicate lanes for each sample. Lanes 1–6: experimental samples, lane 7: no Cas9 control, lane 8: no sgRNA control; ( c ) lane 1: no Cas9 control, lane 2: no sgRNA control, lanes 3–5: experimental samples. Cleavage products at expected band sizes were detected for all samples containing the sgRNA + Cas9 + gDNA. No product at the expected band size was observed for all the control samples.

    Techniques Used: In Vitro, Cleavage Assay, Agarose Gel Electrophoresis

    Ribonucleoprotein (RNP) and plasmid (px458) transfection of ( a ) ASK-1 and ( b ) CHSE-214 cells. Vertical panels show cells transfected with (i) RNP (sgRNA:Cas9-EGFP complex); (ii) Cas9-EGFP protein control; (iii) plasmid (px458); (iv) sham control. Horizontal panels show pictures of cells taken with (1) fluorescent channel; (2) light channel; (3) overlay of the light and fluorescent panels. Pictures were taken at day 7 after RNP electroporation (1600 V, 10 ms, 3 pulses) and day 2 after px458 electroporation (1200 V, 20 ms, 2 pulses). Strong adherence of Cas9-EGFP was observed on the cell surface of CHSE-214, and it was difficult to entirely wash off with PBS, explaining the high background fluorescence in the Cas9-EGFP- and RNP-transfected CHSE-214 cells. Magnification X20. (Fluorescent pictures of transfected SHK-1 cells are not shown).
    Figure Legend Snippet: Ribonucleoprotein (RNP) and plasmid (px458) transfection of ( a ) ASK-1 and ( b ) CHSE-214 cells. Vertical panels show cells transfected with (i) RNP (sgRNA:Cas9-EGFP complex); (ii) Cas9-EGFP protein control; (iii) plasmid (px458); (iv) sham control. Horizontal panels show pictures of cells taken with (1) fluorescent channel; (2) light channel; (3) overlay of the light and fluorescent panels. Pictures were taken at day 7 after RNP electroporation (1600 V, 10 ms, 3 pulses) and day 2 after px458 electroporation (1200 V, 20 ms, 2 pulses). Strong adherence of Cas9-EGFP was observed on the cell surface of CHSE-214, and it was difficult to entirely wash off with PBS, explaining the high background fluorescence in the Cas9-EGFP- and RNP-transfected CHSE-214 cells. Magnification X20. (Fluorescent pictures of transfected SHK-1 cells are not shown).

    Techniques Used: Plasmid Preparation, Transfection, Electroporation, Fluorescence

    Analyses of mutations in the target regions of cr2 and mmp9 genes due to the applied CRISPR/Cas9 strategies. ( a ) T7E1 assay of the cr2 RNP-transfected CHSE-214 cells. Cleaved fragments corresponding to T7 endonuclease 1 digestion of gDNA extract from the RNP-transfected cells are indicated with white arrows in lanes 3 and 4. Similar results were obtained in RNP-transfected ASK-1 and SHK-1 cells for both cr2 and mmp9 . ( b ) Representative Sanger sequencing chromatogram for the target region of cr2 (upper left) and mmp9 (upper right) in ASK-1 cells, either wild-type (WT) or edited with CRISPR/Cas9. The binding regions are represented by the sgRNA rectangular bar, and the edited regions are boxed in dashed lines. ( c ) Representative output of DECODRE analysis of a cr2 RNP-transfected- (lower left) and an mmp9 RNP-transfected (lower right) ASK-1 cell. Indel type and editing frequency are indicated, as well as the aligned sequences of the wild-type and edited samples. The sgRNA including the PAM sequences (red and green bar) highlights the exact target sequence and Cas9 cut site. Boxed dashed lines indicate the exact site of the observed indel mutations.
    Figure Legend Snippet: Analyses of mutations in the target regions of cr2 and mmp9 genes due to the applied CRISPR/Cas9 strategies. ( a ) T7E1 assay of the cr2 RNP-transfected CHSE-214 cells. Cleaved fragments corresponding to T7 endonuclease 1 digestion of gDNA extract from the RNP-transfected cells are indicated with white arrows in lanes 3 and 4. Similar results were obtained in RNP-transfected ASK-1 and SHK-1 cells for both cr2 and mmp9 . ( b ) Representative Sanger sequencing chromatogram for the target region of cr2 (upper left) and mmp9 (upper right) in ASK-1 cells, either wild-type (WT) or edited with CRISPR/Cas9. The binding regions are represented by the sgRNA rectangular bar, and the edited regions are boxed in dashed lines. ( c ) Representative output of DECODRE analysis of a cr2 RNP-transfected- (lower left) and an mmp9 RNP-transfected (lower right) ASK-1 cell. Indel type and editing frequency are indicated, as well as the aligned sequences of the wild-type and edited samples. The sgRNA including the PAM sequences (red and green bar) highlights the exact target sequence and Cas9 cut site. Boxed dashed lines indicate the exact site of the observed indel mutations.

    Techniques Used: CRISPR, Transfection, Sequencing, Binding Assay

    engen sgrna synthesis kit  (New England Biolabs)


    Bioz Verified Symbol New England Biolabs is a verified supplier
    Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    New England Biolabs engen sgrna synthesis kit
    Engen Sgrna Synthesis Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen sgrna synthesis kit/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen sgrna synthesis kit - by Bioz Stars, 2023-02
    86/100 stars

    Images

    engene lbcas12a  (New England Biolabs)


    Bioz Verified Symbol New England Biolabs is a verified supplier
    Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    New England Biolabs engene lbcas12a
    Engene Lbcas12a, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engene lbcas12a/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engene lbcas12a - by Bioz Stars, 2023-02
    86/100 stars

    Images

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86
    New England Biolabs engen sgrna synthesis kit
    Engen Sgrna Synthesis Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen sgrna synthesis kit/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen sgrna synthesis kit - by Bioz Stars, 2023-02
    86/100 stars
      Buy from Supplier

    86
    New England Biolabs engen lba cas12a
    Engen Lba Cas12a, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen lba cas12a/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen lba cas12a - by Bioz Stars, 2023-02
    86/100 stars
      Buy from Supplier

    86
    New England Biolabs engen lba cas12a cpf1 kit
    Feasibility of <t>Cas12a–PMNT-based</t> visual method for nucleic acid detection (1) NEBuffer + positive plasmid DNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (2) NEBuffer + positive plasmid DNA + crRNA + Rnase Inhibitor + ssDNA + PMNT; (3) NEBuffer + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (4) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + PMNT; (5) NEBuffer + ssDNA + PMNT; (6) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (7) NEBuffer + negative plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (8) S1 Reaction Buffer + S1 Nuclease + ssDNA + PMNT.
    Engen Lba Cas12a Cpf1 Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen lba cas12a cpf1 kit/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen lba cas12a cpf1 kit - by Bioz Stars, 2023-02
    86/100 stars
      Buy from Supplier

    86
    New England Biolabs engen sgrna enzyme mix
    Feasibility of <t>Cas12a–PMNT-based</t> visual method for nucleic acid detection (1) NEBuffer + positive plasmid DNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (2) NEBuffer + positive plasmid DNA + crRNA + Rnase Inhibitor + ssDNA + PMNT; (3) NEBuffer + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (4) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + PMNT; (5) NEBuffer + ssDNA + PMNT; (6) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (7) NEBuffer + negative plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (8) S1 Reaction Buffer + S1 Nuclease + ssDNA + PMNT.
    Engen Sgrna Enzyme Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen sgrna enzyme mix/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen sgrna enzyme mix - by Bioz Stars, 2023-02
    86/100 stars
      Buy from Supplier

    86
    New England Biolabs engen 2× sgrna reaction mix
    Single guide RNA sequences and primers used for amplification and sequencing of target genomic regions.
    Engen 2× Sgrna Reaction Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engen 2× sgrna reaction mix/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engen 2× sgrna reaction mix - by Bioz Stars, 2023-02
    86/100 stars
      Buy from Supplier

    86
    New England Biolabs engene lbcas12a
    Single guide RNA sequences and primers used for amplification and sequencing of target genomic regions.
    Engene Lbcas12a, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/engene lbcas12a/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    engene lbcas12a - by Bioz Stars, 2023-02
    86/100 stars
      Buy from Supplier

    Image Search Results


    Feasibility of Cas12a–PMNT-based visual method for nucleic acid detection (1) NEBuffer + positive plasmid DNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (2) NEBuffer + positive plasmid DNA + crRNA + Rnase Inhibitor + ssDNA + PMNT; (3) NEBuffer + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (4) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + PMNT; (5) NEBuffer + ssDNA + PMNT; (6) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (7) NEBuffer + negative plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (8) S1 Reaction Buffer + S1 Nuclease + ssDNA + PMNT.

    Journal: Foods

    Article Title: A Rapid and Visual Method for Nucleic Acid Detection of Escherichia coli O157:H7 Based on CRISPR/Cas12a-PMNT

    doi: 10.3390/foods12020236

    Figure Lengend Snippet: Feasibility of Cas12a–PMNT-based visual method for nucleic acid detection (1) NEBuffer + positive plasmid DNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (2) NEBuffer + positive plasmid DNA + crRNA + Rnase Inhibitor + ssDNA + PMNT; (3) NEBuffer + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (4) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + PMNT; (5) NEBuffer + ssDNA + PMNT; (6) NEBuffer + positive plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (7) NEBuffer + negative plasmid DNA + crRNA + Cas12a + Rnase Inhibitor + ssDNA + PMNT; (8) S1 Reaction Buffer + S1 Nuclease + ssDNA + PMNT.

    Article Snippet: EnGen Lba Cas12a (Cpf1) kit for Cas12a-mediated target cleavage assay was purchased from New England Biolabs Inc. (Ipswich, MA, USA).

    Techniques: Plasmid Preparation

    Single guide RNA sequences and primers used for amplification and sequencing of target genomic regions.

    Journal: International Journal of Molecular Sciences

    Article Title: CRISPR/Cas9-Mediated Gene Editing in Salmonids Cells and Efficient Establishment of Edited Clonal Cell Lines

    doi: 10.3390/ijms232416218

    Figure Lengend Snippet: Single guide RNA sequences and primers used for amplification and sequencing of target genomic regions.

    Article Snippet: The laboratory synthesis of the sgRNA was carried out by mixing the target-specific oligos with the manufacturer’s (New England Biolabs, Hitchin, UK) reaction mix, comprising nuclease-free water (2 μL); EnGen 2× sgRNA reaction mix (dNTPs, S. pyogenes Cas9 scaffold oligo) (10 μL); target-specific DNA oligo (1 μM; 5 μL); DTT (0.1 M; 1 μL); and EnGen sgRNA Enzyme mix (2 μL).

    Techniques: Amplification, Sequencing

    sgRNA analysis and in vitro cleavage assay for cr2 and mmp9 genes. ( a ) sgRNA analysis on 2% agarose gel. UD: undenatured sgRNA, D: denatured sgRNA. Single bands detected for the denatured sgRNAs highlighted the purity of the products, and multiple bands in the undenatured sgRNA resulted from the presence of RNA secondary structures; ( b ) gel shows duplicate lanes for each sample. Lanes 1–6: experimental samples, lane 7: no Cas9 control, lane 8: no sgRNA control; ( c ) lane 1: no Cas9 control, lane 2: no sgRNA control, lanes 3–5: experimental samples. Cleavage products at expected band sizes were detected for all samples containing the sgRNA + Cas9 + gDNA. No product at the expected band size was observed for all the control samples.

    Journal: International Journal of Molecular Sciences

    Article Title: CRISPR/Cas9-Mediated Gene Editing in Salmonids Cells and Efficient Establishment of Edited Clonal Cell Lines

    doi: 10.3390/ijms232416218

    Figure Lengend Snippet: sgRNA analysis and in vitro cleavage assay for cr2 and mmp9 genes. ( a ) sgRNA analysis on 2% agarose gel. UD: undenatured sgRNA, D: denatured sgRNA. Single bands detected for the denatured sgRNAs highlighted the purity of the products, and multiple bands in the undenatured sgRNA resulted from the presence of RNA secondary structures; ( b ) gel shows duplicate lanes for each sample. Lanes 1–6: experimental samples, lane 7: no Cas9 control, lane 8: no sgRNA control; ( c ) lane 1: no Cas9 control, lane 2: no sgRNA control, lanes 3–5: experimental samples. Cleavage products at expected band sizes were detected for all samples containing the sgRNA + Cas9 + gDNA. No product at the expected band size was observed for all the control samples.

    Article Snippet: The laboratory synthesis of the sgRNA was carried out by mixing the target-specific oligos with the manufacturer’s (New England Biolabs, Hitchin, UK) reaction mix, comprising nuclease-free water (2 μL); EnGen 2× sgRNA reaction mix (dNTPs, S. pyogenes Cas9 scaffold oligo) (10 μL); target-specific DNA oligo (1 μM; 5 μL); DTT (0.1 M; 1 μL); and EnGen sgRNA Enzyme mix (2 μL).

    Techniques: In Vitro, Cleavage Assay, Agarose Gel Electrophoresis

    Ribonucleoprotein (RNP) and plasmid (px458) transfection of ( a ) ASK-1 and ( b ) CHSE-214 cells. Vertical panels show cells transfected with (i) RNP (sgRNA:Cas9-EGFP complex); (ii) Cas9-EGFP protein control; (iii) plasmid (px458); (iv) sham control. Horizontal panels show pictures of cells taken with (1) fluorescent channel; (2) light channel; (3) overlay of the light and fluorescent panels. Pictures were taken at day 7 after RNP electroporation (1600 V, 10 ms, 3 pulses) and day 2 after px458 electroporation (1200 V, 20 ms, 2 pulses). Strong adherence of Cas9-EGFP was observed on the cell surface of CHSE-214, and it was difficult to entirely wash off with PBS, explaining the high background fluorescence in the Cas9-EGFP- and RNP-transfected CHSE-214 cells. Magnification X20. (Fluorescent pictures of transfected SHK-1 cells are not shown).

    Journal: International Journal of Molecular Sciences

    Article Title: CRISPR/Cas9-Mediated Gene Editing in Salmonids Cells and Efficient Establishment of Edited Clonal Cell Lines

    doi: 10.3390/ijms232416218

    Figure Lengend Snippet: Ribonucleoprotein (RNP) and plasmid (px458) transfection of ( a ) ASK-1 and ( b ) CHSE-214 cells. Vertical panels show cells transfected with (i) RNP (sgRNA:Cas9-EGFP complex); (ii) Cas9-EGFP protein control; (iii) plasmid (px458); (iv) sham control. Horizontal panels show pictures of cells taken with (1) fluorescent channel; (2) light channel; (3) overlay of the light and fluorescent panels. Pictures were taken at day 7 after RNP electroporation (1600 V, 10 ms, 3 pulses) and day 2 after px458 electroporation (1200 V, 20 ms, 2 pulses). Strong adherence of Cas9-EGFP was observed on the cell surface of CHSE-214, and it was difficult to entirely wash off with PBS, explaining the high background fluorescence in the Cas9-EGFP- and RNP-transfected CHSE-214 cells. Magnification X20. (Fluorescent pictures of transfected SHK-1 cells are not shown).

    Article Snippet: The laboratory synthesis of the sgRNA was carried out by mixing the target-specific oligos with the manufacturer’s (New England Biolabs, Hitchin, UK) reaction mix, comprising nuclease-free water (2 μL); EnGen 2× sgRNA reaction mix (dNTPs, S. pyogenes Cas9 scaffold oligo) (10 μL); target-specific DNA oligo (1 μM; 5 μL); DTT (0.1 M; 1 μL); and EnGen sgRNA Enzyme mix (2 μL).

    Techniques: Plasmid Preparation, Transfection, Electroporation, Fluorescence

    Analyses of mutations in the target regions of cr2 and mmp9 genes due to the applied CRISPR/Cas9 strategies. ( a ) T7E1 assay of the cr2 RNP-transfected CHSE-214 cells. Cleaved fragments corresponding to T7 endonuclease 1 digestion of gDNA extract from the RNP-transfected cells are indicated with white arrows in lanes 3 and 4. Similar results were obtained in RNP-transfected ASK-1 and SHK-1 cells for both cr2 and mmp9 . ( b ) Representative Sanger sequencing chromatogram for the target region of cr2 (upper left) and mmp9 (upper right) in ASK-1 cells, either wild-type (WT) or edited with CRISPR/Cas9. The binding regions are represented by the sgRNA rectangular bar, and the edited regions are boxed in dashed lines. ( c ) Representative output of DECODRE analysis of a cr2 RNP-transfected- (lower left) and an mmp9 RNP-transfected (lower right) ASK-1 cell. Indel type and editing frequency are indicated, as well as the aligned sequences of the wild-type and edited samples. The sgRNA including the PAM sequences (red and green bar) highlights the exact target sequence and Cas9 cut site. Boxed dashed lines indicate the exact site of the observed indel mutations.

    Journal: International Journal of Molecular Sciences

    Article Title: CRISPR/Cas9-Mediated Gene Editing in Salmonids Cells and Efficient Establishment of Edited Clonal Cell Lines

    doi: 10.3390/ijms232416218

    Figure Lengend Snippet: Analyses of mutations in the target regions of cr2 and mmp9 genes due to the applied CRISPR/Cas9 strategies. ( a ) T7E1 assay of the cr2 RNP-transfected CHSE-214 cells. Cleaved fragments corresponding to T7 endonuclease 1 digestion of gDNA extract from the RNP-transfected cells are indicated with white arrows in lanes 3 and 4. Similar results were obtained in RNP-transfected ASK-1 and SHK-1 cells for both cr2 and mmp9 . ( b ) Representative Sanger sequencing chromatogram for the target region of cr2 (upper left) and mmp9 (upper right) in ASK-1 cells, either wild-type (WT) or edited with CRISPR/Cas9. The binding regions are represented by the sgRNA rectangular bar, and the edited regions are boxed in dashed lines. ( c ) Representative output of DECODRE analysis of a cr2 RNP-transfected- (lower left) and an mmp9 RNP-transfected (lower right) ASK-1 cell. Indel type and editing frequency are indicated, as well as the aligned sequences of the wild-type and edited samples. The sgRNA including the PAM sequences (red and green bar) highlights the exact target sequence and Cas9 cut site. Boxed dashed lines indicate the exact site of the observed indel mutations.

    Article Snippet: The laboratory synthesis of the sgRNA was carried out by mixing the target-specific oligos with the manufacturer’s (New England Biolabs, Hitchin, UK) reaction mix, comprising nuclease-free water (2 μL); EnGen 2× sgRNA reaction mix (dNTPs, S. pyogenes Cas9 scaffold oligo) (10 μL); target-specific DNA oligo (1 μM; 5 μL); DTT (0.1 M; 1 μL); and EnGen sgRNA Enzyme mix (2 μL).

    Techniques: CRISPR, Transfection, Sequencing, Binding Assay