haeiii  (New England Biolabs)


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    Structured Review

    New England Biolabs haeiii
    NHEJ-mediated integration of MRSs in HEK293T cells. ( a ) Gel image showing T7EI cleavage products (red brackets) of edited BCL11A-XL 3′ UTR 1 site. ( b ) Quantification of T7EI gel bands showing similar percentages of genome editing across samples (32.6% to 44.8%). ( c ) Apparent absence of PCR-RFLP <t>HaeIII</t> cleavage for test samples, indicative of dsODN integration being below the detection threshold. ( d ) Top: schematic illustration of dsODN-specific amplification assay at the 3′ UTR 1 site; bottom: amplicons of ~250 and ~302 bp revealing the presence of 2 MRSs and 4 MRSs in cells, respectively. Of note, undesirable inverted integration of MRSs into 3′ UTR 1 (see greyed-out part of Figure 2 ) is not detected by this assay. MOCK: mock-nucleofected cell sample, NO DONOR: cell sample nucleofected only with RNPs, MRS: miRNA recognition site, <t>dsODN451-2MRSs/*:</t> cell samples nucleofected with RNPs and dsODN bearing two miR451a MRSs at the indicated picomole quantity, dsODN451-4MRSs/5: cell sample nucleofected with RNPs and 5 pmole dsODN bearing four miR451a MRSs, +: a 994-bp PCR product giving cleavage products of 572 and 422 bp after digestion with HaeIII.
    Haeiii, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/haeiii/product/New England Biolabs
    Average 97 stars, based on 40 article reviews
    Price from $9.99 to $1999.99
    haeiii - by Bioz Stars, 2022-12
    97/100 stars

    Images

    1) Product Images from "CRISPR Editing Enables Consequential Tag-Activated MicroRNA-Mediated Endogene Deactivation"

    Article Title: CRISPR Editing Enables Consequential Tag-Activated MicroRNA-Mediated Endogene Deactivation

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms23031082

    NHEJ-mediated integration of MRSs in HEK293T cells. ( a ) Gel image showing T7EI cleavage products (red brackets) of edited BCL11A-XL 3′ UTR 1 site. ( b ) Quantification of T7EI gel bands showing similar percentages of genome editing across samples (32.6% to 44.8%). ( c ) Apparent absence of PCR-RFLP HaeIII cleavage for test samples, indicative of dsODN integration being below the detection threshold. ( d ) Top: schematic illustration of dsODN-specific amplification assay at the 3′ UTR 1 site; bottom: amplicons of ~250 and ~302 bp revealing the presence of 2 MRSs and 4 MRSs in cells, respectively. Of note, undesirable inverted integration of MRSs into 3′ UTR 1 (see greyed-out part of Figure 2 ) is not detected by this assay. MOCK: mock-nucleofected cell sample, NO DONOR: cell sample nucleofected only with RNPs, MRS: miRNA recognition site, dsODN451-2MRSs/*: cell samples nucleofected with RNPs and dsODN bearing two miR451a MRSs at the indicated picomole quantity, dsODN451-4MRSs/5: cell sample nucleofected with RNPs and 5 pmole dsODN bearing four miR451a MRSs, +: a 994-bp PCR product giving cleavage products of 572 and 422 bp after digestion with HaeIII.
    Figure Legend Snippet: NHEJ-mediated integration of MRSs in HEK293T cells. ( a ) Gel image showing T7EI cleavage products (red brackets) of edited BCL11A-XL 3′ UTR 1 site. ( b ) Quantification of T7EI gel bands showing similar percentages of genome editing across samples (32.6% to 44.8%). ( c ) Apparent absence of PCR-RFLP HaeIII cleavage for test samples, indicative of dsODN integration being below the detection threshold. ( d ) Top: schematic illustration of dsODN-specific amplification assay at the 3′ UTR 1 site; bottom: amplicons of ~250 and ~302 bp revealing the presence of 2 MRSs and 4 MRSs in cells, respectively. Of note, undesirable inverted integration of MRSs into 3′ UTR 1 (see greyed-out part of Figure 2 ) is not detected by this assay. MOCK: mock-nucleofected cell sample, NO DONOR: cell sample nucleofected only with RNPs, MRS: miRNA recognition site, dsODN451-2MRSs/*: cell samples nucleofected with RNPs and dsODN bearing two miR451a MRSs at the indicated picomole quantity, dsODN451-4MRSs/5: cell sample nucleofected with RNPs and 5 pmole dsODN bearing four miR451a MRSs, +: a 994-bp PCR product giving cleavage products of 572 and 422 bp after digestion with HaeIII.

    Techniques Used: Non-Homologous End Joining, Polymerase Chain Reaction, Amplification

    HDR-mediated integration of MRSs in HUDEP-2 cells. PCR-RFLP analysis of cells nucleofected with RNPs and ss DNA oligos was performed for the assessment of integration efficiency. Bands of 436 bp represent unmodified target sites, larger bands insertions with incomplete digestion by DdeI/HaeIII, and smaller bands cleavage products of insertions. All bands corresponding to insertions are indicated by arrowheads (band sizes of 554 bp and 277 bp for donors with 2 MRSs, and of 528 bp and 264 bp for donors with 1 MRS). Corresponding rates of DdeI/HaeIII cleavage in PCR-RFLP (%) (after subtraction of background average cleavage rates of control samples) are reported below the gels. ( a ) Analysis of cells nucleofected with RNPs and Ultramer DNA Oligos. ( b ) Analysis of cells nucleofected with Alt-R RNPs and Alt-R HDR-2MRSs DNA Oligo (left gel), shown in comparison with analysis of cells nucleofected with standard Cas9 RNPs and Ultramer ssODN451TS_2MRSs DNA Oligo (right gel). Separate gels are indicated by dashed lines. MOCK: mock-nucleofected sample, NO DONOR: cell sample nucleofected only with RNPs, MRS: miRNA recognition site, NTS: non-target strand, TS: target strand, +: PCR products bearing restriction sites for DdeI/HaeIII used as positive controls. See Section 4.2 and Supplementary Table S2 for ss DNA oligo naming.
    Figure Legend Snippet: HDR-mediated integration of MRSs in HUDEP-2 cells. PCR-RFLP analysis of cells nucleofected with RNPs and ss DNA oligos was performed for the assessment of integration efficiency. Bands of 436 bp represent unmodified target sites, larger bands insertions with incomplete digestion by DdeI/HaeIII, and smaller bands cleavage products of insertions. All bands corresponding to insertions are indicated by arrowheads (band sizes of 554 bp and 277 bp for donors with 2 MRSs, and of 528 bp and 264 bp for donors with 1 MRS). Corresponding rates of DdeI/HaeIII cleavage in PCR-RFLP (%) (after subtraction of background average cleavage rates of control samples) are reported below the gels. ( a ) Analysis of cells nucleofected with RNPs and Ultramer DNA Oligos. ( b ) Analysis of cells nucleofected with Alt-R RNPs and Alt-R HDR-2MRSs DNA Oligo (left gel), shown in comparison with analysis of cells nucleofected with standard Cas9 RNPs and Ultramer ssODN451TS_2MRSs DNA Oligo (right gel). Separate gels are indicated by dashed lines. MOCK: mock-nucleofected sample, NO DONOR: cell sample nucleofected only with RNPs, MRS: miRNA recognition site, NTS: non-target strand, TS: target strand, +: PCR products bearing restriction sites for DdeI/HaeIII used as positive controls. See Section 4.2 and Supplementary Table S2 for ss DNA oligo naming.

    Techniques Used: Polymerase Chain Reaction

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    New England Biolabs hae iii
    Effect of some physicochemical parameters and different concentrations of As(V) and <t>As(III)</t> on bacterial growth. ( A ) Temperature, ( B ) pH, ( C ) NaCl, ( D ) As(V), ( E ) As(III).
    Hae Iii, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hae iii/product/New England Biolabs
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hae iii - by Bioz Stars, 2022-12
    96/100 stars
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    Effect of some physicochemical parameters and different concentrations of As(V) and As(III) on bacterial growth. ( A ) Temperature, ( B ) pH, ( C ) NaCl, ( D ) As(V), ( E ) As(III).

    Journal: Life

    Article Title: Arsenic Pollution and Anaerobic Arsenic Metabolizing Bacteria in Lake Van, the World’s Largest Soda Lake

    doi: 10.3390/life12111900

    Figure Lengend Snippet: Effect of some physicochemical parameters and different concentrations of As(V) and As(III) on bacterial growth. ( A ) Temperature, ( B ) pH, ( C ) NaCl, ( D ) As(V), ( E ) As(III).

    Article Snippet: To reveal individual differences, amplified 16S rRNA gene regions of each isolate were cut with Eco RI (CutSmart Biolabs, New England Biolabs, Ipswich, MA, USA, R3101S), Rsa I (CutSmart Biolabs, New England Biolabs, R0167S), and Hae III (CutSmart Biolabs, New England Biolabs, R01108S) restriction enzymes according to manufacturer’s instructions.

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