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    Thermo Fisher exonuclease i thermo fisher scientific
    Principle of mismatch removal. In the first step, a double-stranded polynucleotide with a mismatching base pair, e.g. resulting from ligation of oligonucleotides in a gene synthesis reaction, is cleaved by the EMC enzyme in both strands, 2–5 bp downstream of the mismatch. The short overhangs thus generated dissociate immediately at the reaction temperature of 37°C. The resulting single-stranded overhangs can be removed by different strategies: addition of a single-strand-specific 3′–5′-exonuclease, e.g. E.coli <t>exonuclease</t> I in the EMC reaction or in a subsequent exonuclease step. Alternatively, the 3′–5′-exonuclease activity of proofreading polymerases can be used in the subsequent PCR. The proposed mechanism in this case is (i) removal of single-stranded overhangs during the initial heating step from 20 to 95°C or (ii) removal of mispaired bases by ‘proofreading’ in the first elongation cycle of the overlap extension PCR.
    Exonuclease I Thermo Fisher Scientific, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 146 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/exonuclease i thermo fisher scientific/product/Thermo Fisher
    Average 99 stars, based on 146 article reviews
    Price from $9.99 to $1999.99
    exonuclease i thermo fisher scientific - by Bioz Stars, 2020-08
    99/100 stars
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    85
    Thermo Fisher exonuclease 1 exo1
    Principle of mismatch removal. In the first step, a double-stranded polynucleotide with a mismatching base pair, e.g. resulting from ligation of oligonucleotides in a gene synthesis reaction, is cleaved by the EMC enzyme in both strands, 2–5 bp downstream of the mismatch. The short overhangs thus generated dissociate immediately at the reaction temperature of 37°C. The resulting single-stranded overhangs can be removed by different strategies: addition of a single-strand-specific 3′–5′-exonuclease, e.g. E.coli <t>exonuclease</t> I in the EMC reaction or in a subsequent exonuclease step. Alternatively, the 3′–5′-exonuclease activity of proofreading polymerases can be used in the subsequent PCR. The proposed mechanism in this case is (i) removal of single-stranded overhangs during the initial heating step from 20 to 95°C or (ii) removal of mispaired bases by ‘proofreading’ in the first elongation cycle of the overlap extension PCR.
    Exonuclease 1 Exo1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/exonuclease 1 exo1/product/Thermo Fisher
    Average 85 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    exonuclease 1 exo1 - by Bioz Stars, 2020-08
    85/100 stars
      Buy from Supplier

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    Principle of mismatch removal. In the first step, a double-stranded polynucleotide with a mismatching base pair, e.g. resulting from ligation of oligonucleotides in a gene synthesis reaction, is cleaved by the EMC enzyme in both strands, 2–5 bp downstream of the mismatch. The short overhangs thus generated dissociate immediately at the reaction temperature of 37°C. The resulting single-stranded overhangs can be removed by different strategies: addition of a single-strand-specific 3′–5′-exonuclease, e.g. E.coli exonuclease I in the EMC reaction or in a subsequent exonuclease step. Alternatively, the 3′–5′-exonuclease activity of proofreading polymerases can be used in the subsequent PCR. The proposed mechanism in this case is (i) removal of single-stranded overhangs during the initial heating step from 20 to 95°C or (ii) removal of mispaired bases by ‘proofreading’ in the first elongation cycle of the overlap extension PCR.

    Journal: Nucleic Acids Research

    Article Title: Removal of mismatched bases from synthetic genes by enzymatic mismatch cleavage

    doi: 10.1093/nar/gni058

    Figure Lengend Snippet: Principle of mismatch removal. In the first step, a double-stranded polynucleotide with a mismatching base pair, e.g. resulting from ligation of oligonucleotides in a gene synthesis reaction, is cleaved by the EMC enzyme in both strands, 2–5 bp downstream of the mismatch. The short overhangs thus generated dissociate immediately at the reaction temperature of 37°C. The resulting single-stranded overhangs can be removed by different strategies: addition of a single-strand-specific 3′–5′-exonuclease, e.g. E.coli exonuclease I in the EMC reaction or in a subsequent exonuclease step. Alternatively, the 3′–5′-exonuclease activity of proofreading polymerases can be used in the subsequent PCR. The proposed mechanism in this case is (i) removal of single-stranded overhangs during the initial heating step from 20 to 95°C or (ii) removal of mispaired bases by ‘proofreading’ in the first elongation cycle of the overlap extension PCR.

    Article Snippet: Cleavage was performed at 37°C in the buffer recommended by the manufacturer, for up to 24 h. For removal of single-stranded DNA, E.coli exonuclease I (Fermentas, St Leon-Rot, Germany) was included in some reactions.

    Techniques: Ligation, Generated, Activity Assay, Polymerase Chain Reaction

    Alkaline denaturation method. (A) Experimental design. (B) lane A – emulsion PCR product (step2), lane B – Nb.BtsI nicked ssDNA (step 3), lane C – nicked DNA (step 4), lane D alkali-melting of nicked ssDNA (step 6), lane E – one of negative selection (step 7), lane F – exonuclease I hydrolysis step 7 products.

    Journal: PLoS ONE

    Article Title: Methods for the Preparation of Large Quantities of Complex Single-Stranded Oligonucleotide Libraries

    doi: 10.1371/journal.pone.0094752

    Figure Lengend Snippet: Alkaline denaturation method. (A) Experimental design. (B) lane A – emulsion PCR product (step2), lane B – Nb.BtsI nicked ssDNA (step 3), lane C – nicked DNA (step 4), lane D alkali-melting of nicked ssDNA (step 6), lane E – one of negative selection (step 7), lane F – exonuclease I hydrolysis step 7 products.

    Article Snippet: DNA exonuclease I and Antartic phosphatase are obtained from Fermentas.

    Techniques: Polymerase Chain Reaction, Selection

    Mutations in STN1 result in abnormal telomere phenotypes. (A) DNA samples, prepared from PBLs of patient P1, her heterozygous father (F1), and a noncarrier sibling (S1) and patient P2, his heterozygous mother (M2), and two independent control samples (C), were analyzed by in-gel hybridization. Duplicated lanes were electrophoresed in the same gel, and then separated and hybridized to a G-rich or C-rich telomeric probe, as indicated above the panels. After native hybridization to detect single-stranded telomeric DNA (top), the gels were denatured and rehybridized with the same probes to detect the overall duplex telomeric DNA (bottom). Treatment with exonuclease I is indicated above the lanes. (B) Graphic illustration of the mean telomere length for the patients and their family members, calculated based on the following number of independent measurements of four in-gels and two Southern analyses: P1:6, M1:3, F1:3, S1:3, P2:9, M2:3, F2:1, C1:2, and C2:3. (C) Graphic illustration of the relative native (single strand) per denatured (total) telomeric signal, normalized to the controls. The values represent the mean of four independent measurements for P1 and four for P2. (D, top) A metaphase spread from P2 PBL after CO-FISH. Bar, 5 µm. The area designated by the white frame in enlarged in the bottom panel. Arrowheads point to chromosome ends with hybridization signals on both sister chromatids, indicating that T-SCE occurred at that chromosome end. (right) The frequency of T-SCE in P2 PBLs is compared with PBLs from four age-matched controls (con-BL1-4). For P2 PBLs, 1,150 chromosome ends were analyzed. For control PBLs, between 600 and 930 telomeres were analyzed (*, P

    Journal: The Journal of Experimental Medicine

    Article Title: Mutations in STN1 cause Coats plus syndrome and are associated with genomic and telomere defects

    doi: 10.1084/jem.20151618

    Figure Lengend Snippet: Mutations in STN1 result in abnormal telomere phenotypes. (A) DNA samples, prepared from PBLs of patient P1, her heterozygous father (F1), and a noncarrier sibling (S1) and patient P2, his heterozygous mother (M2), and two independent control samples (C), were analyzed by in-gel hybridization. Duplicated lanes were electrophoresed in the same gel, and then separated and hybridized to a G-rich or C-rich telomeric probe, as indicated above the panels. After native hybridization to detect single-stranded telomeric DNA (top), the gels were denatured and rehybridized with the same probes to detect the overall duplex telomeric DNA (bottom). Treatment with exonuclease I is indicated above the lanes. (B) Graphic illustration of the mean telomere length for the patients and their family members, calculated based on the following number of independent measurements of four in-gels and two Southern analyses: P1:6, M1:3, F1:3, S1:3, P2:9, M2:3, F2:1, C1:2, and C2:3. (C) Graphic illustration of the relative native (single strand) per denatured (total) telomeric signal, normalized to the controls. The values represent the mean of four independent measurements for P1 and four for P2. (D, top) A metaphase spread from P2 PBL after CO-FISH. Bar, 5 µm. The area designated by the white frame in enlarged in the bottom panel. Arrowheads point to chromosome ends with hybridization signals on both sister chromatids, indicating that T-SCE occurred at that chromosome end. (right) The frequency of T-SCE in P2 PBLs is compared with PBLs from four age-matched controls (con-BL1-4). For P2 PBLs, 1,150 chromosome ends were analyzed. For control PBLs, between 600 and 930 telomeres were analyzed (*, P

    Article Snippet: Exonuclease I treatment was performed overnight at 37°C with 200 U exonuclease I (Thermo Fisher Scientific) per 5 µg DNA in 500 µl, followed by ethanol precipitation.

    Techniques: Hybridization, Fluorescence In Situ Hybridization