homology directed repair kpn i hf  (New England Biolabs)


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    Name:
    KpnI
    Description:
    KpnI 20 000 units
    Catalog Number:
    R0142L
    Price:
    269
    Category:
    Restriction Enzymes
    Size:
    20 000 units
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    Structured Review

    New England Biolabs homology directed repair kpn i hf
    KpnI
    KpnI 20 000 units
    https://www.bioz.com/result/homology directed repair kpn i hf/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    homology directed repair kpn i hf - by Bioz Stars, 2021-06
    99/100 stars

    Images

    1) Product Images from "Effective CRISPR/Cas9-mediated correction of a Fanconi anemia defect by error-prone end joining or templated repair"

    Article Title: Effective CRISPR/Cas9-mediated correction of a Fanconi anemia defect by error-prone end joining or templated repair

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-36506-w

    CRISPR/Cas9 templated gene editing analysis in mouse fibroblasts. ( A ) Overview of the sgRNA + 1 target site with sections of the sense and antisense (AS) ssODN that remove the c.828TAAA insertion and introduce a Kpn I recognition site by a silent C > A substitution. Predicted cut site for Cas9 are indicated by “ ˄ ” and “ ”, with “ ” indicating the strand that is nicked by Cas9D10A. Nucleotide numbers refer to the mutant c.828InsTAAA Fancf gene. ( B ) Overall gene editing efficiencies obtained in mouse fibroblasts depicted as general indel frequencies determined by TIDER (n = 3, one sided Student’s T-test, *p = 0.02). ( C ) Restriction fragment length polymorphism (RFLP) analysis by Kpn I digest of the Fancf locus provides evidence for template-based repair after Cas9, sgRNA + 1 and ssODN transfection (n = 3, one sided Student’s T-test, *p
    Figure Legend Snippet: CRISPR/Cas9 templated gene editing analysis in mouse fibroblasts. ( A ) Overview of the sgRNA + 1 target site with sections of the sense and antisense (AS) ssODN that remove the c.828TAAA insertion and introduce a Kpn I recognition site by a silent C > A substitution. Predicted cut site for Cas9 are indicated by “ ˄ ” and “ ”, with “ ” indicating the strand that is nicked by Cas9D10A. Nucleotide numbers refer to the mutant c.828InsTAAA Fancf gene. ( B ) Overall gene editing efficiencies obtained in mouse fibroblasts depicted as general indel frequencies determined by TIDER (n = 3, one sided Student’s T-test, *p = 0.02). ( C ) Restriction fragment length polymorphism (RFLP) analysis by Kpn I digest of the Fancf locus provides evidence for template-based repair after Cas9, sgRNA + 1 and ssODN transfection (n = 3, one sided Student’s T-test, *p

    Techniques Used: CRISPR, Introduce, Mutagenesis, Transfection

    CRISPR/Cas9 templated gene editing analysis in mouse ES cells. ( A ) Overall gene editing efficiencies obtained in mouse embryonic stem cells depicted as general indel frequencies determined by TIDER (n = 3, one sided Student’s T-test, *p ≤ 0.02). ( B ) TIDE analysis of indel formation at predicted off-target loci after wtCas9 and sgRNA + 1 exposure irrespective of the ssODN orientation (n = 3, one sided Student’s T-test, *p ≤ 0.009). ( C ) Restriction fragment length polymorphism (RFLP) analysis by Kpn I digest of the Fancf locus provides evidence for template-based repair after Cas9 transfection (n = 6, one sided Student’s T-test, *p ≤ 0.02). ( D ) Mouse ES cells show enhanced clonal survival in the presence of 12.5 nM MMC after double-strand break formation by Cas9 and sgRNA + 1 independent of the orientation of the HDR template. A significant difference in relative mESC clone survival was also observed with the antisense (AS) ssODN following Cas9 nickase activity (n = 6, one sided Student’s T-test, *p ≤ 0.025), “M” = mock sgRNA.
    Figure Legend Snippet: CRISPR/Cas9 templated gene editing analysis in mouse ES cells. ( A ) Overall gene editing efficiencies obtained in mouse embryonic stem cells depicted as general indel frequencies determined by TIDER (n = 3, one sided Student’s T-test, *p ≤ 0.02). ( B ) TIDE analysis of indel formation at predicted off-target loci after wtCas9 and sgRNA + 1 exposure irrespective of the ssODN orientation (n = 3, one sided Student’s T-test, *p ≤ 0.009). ( C ) Restriction fragment length polymorphism (RFLP) analysis by Kpn I digest of the Fancf locus provides evidence for template-based repair after Cas9 transfection (n = 6, one sided Student’s T-test, *p ≤ 0.02). ( D ) Mouse ES cells show enhanced clonal survival in the presence of 12.5 nM MMC after double-strand break formation by Cas9 and sgRNA + 1 independent of the orientation of the HDR template. A significant difference in relative mESC clone survival was also observed with the antisense (AS) ssODN following Cas9 nickase activity (n = 6, one sided Student’s T-test, *p ≤ 0.025), “M” = mock sgRNA.

    Techniques Used: CRISPR, Transfection, Activity Assay

    Related Articles

    Southern Blot:

    Article Title: Molecular analysis of a locus control region in the T helper 2 cytokine gene cluster: A target for STAT6 but not GATA3
    Article Snippet: .. Overlapping Bam HI and Kpn I (enzymes from New England Biolabs, Beverly, MA) genomic fragments spanning KIF3A to RAD50 were chosen for Southern blot analysis. ..

    Plasmid Preparation:

    Article Title: Recognition of DNA Termini by the C-Terminal Region of the Ku80 and the DNA-Dependent Protein Kinase Catalytic Subunit
    Article Snippet: .. Kinase assays using plasmid DNA substrates were performed with pcDNA3.1 digested with either XhoI, BamHI, EcoRV, and KpnI or pCAG-GFP digested with XbaI or EcoRI (New England Biolabs). .. The specific sequences recognized by the restriction enzymes and DNA termini generated are shown in .

    Article Title: Repurposing the Native Type I-F CRISPR-Cas System in Pseudomonas aeruginosa for Genome Editing
    Article Snippet: Alternatives: If the Kpn I-mini-CRISPR-BamHI fragment is synthesized and supplied in the linear double-stranded DNA form, it can be directly proceeded to Step 3. .. Purified PCR product from Step 2 and the plasmid pAY5211 are digested with Kpn I and BamHI (NEB, USA). ..

    Binding Assay:

    Article Title: RNA aptamer inhibitors of a restriction endonuclease
    Article Snippet: .. Binding reactions contain radiolabeled aptamer (13 nM) and the indicated concentration of KpnI in New England Biolabs 1.1 buffer (10 mM Bis-Tris-Propane-HCl, pH 7, 10 mM MgCl2 , 100 μg/ml bovine serum albumin (BSA)) for 30 min at 37°C. ..

    Concentration Assay:

    Article Title: RNA aptamer inhibitors of a restriction endonuclease
    Article Snippet: .. Binding reactions contain radiolabeled aptamer (13 nM) and the indicated concentration of KpnI in New England Biolabs 1.1 buffer (10 mM Bis-Tris-Propane-HCl, pH 7, 10 mM MgCl2 , 100 μg/ml bovine serum albumin (BSA)) for 30 min at 37°C. ..

    Purification:

    Article Title: Repurposing the Native Type I-F CRISPR-Cas System in Pseudomonas aeruginosa for Genome Editing
    Article Snippet: Alternatives: If the Kpn I-mini-CRISPR-BamHI fragment is synthesized and supplied in the linear double-stranded DNA form, it can be directly proceeded to Step 3. .. Purified PCR product from Step 2 and the plasmid pAY5211 are digested with Kpn I and BamHI (NEB, USA). ..

    Polymerase Chain Reaction:

    Article Title: Repurposing the Native Type I-F CRISPR-Cas System in Pseudomonas aeruginosa for Genome Editing
    Article Snippet: Alternatives: If the Kpn I-mini-CRISPR-BamHI fragment is synthesized and supplied in the linear double-stranded DNA form, it can be directly proceeded to Step 3. .. Purified PCR product from Step 2 and the plasmid pAY5211 are digested with Kpn I and BamHI (NEB, USA). ..

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    New England Biolabs homology directed repair kpn i hf
    CRISPR/Cas9 templated gene editing analysis in mouse fibroblasts. ( A ) Overview of the sgRNA + 1 target site with sections of the sense and antisense (AS) <t>ssODN</t> that remove the c.828TAAA insertion and introduce a <t>Kpn</t> I recognition site by a silent C > A substitution. Predicted cut site for Cas9 are indicated by “ ˄ ” and “ ”, with “ ” indicating the strand that is nicked by Cas9D10A. Nucleotide numbers refer to the mutant c.828InsTAAA Fancf gene. ( B ) Overall gene editing efficiencies obtained in mouse fibroblasts depicted as general indel frequencies determined by TIDER (n = 3, one sided Student’s T-test, *p = 0.02). ( C ) Restriction fragment length polymorphism (RFLP) analysis by Kpn I digest of the Fancf locus provides evidence for template-based repair after Cas9, sgRNA + 1 and ssODN transfection (n = 3, one sided Student’s T-test, *p
    Homology Directed Repair Kpn I Hf, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/homology directed repair kpn i hf/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    homology directed repair kpn i hf - by Bioz Stars, 2021-06
    99/100 stars
      Buy from Supplier

    Image Search Results


    CRISPR/Cas9 templated gene editing analysis in mouse fibroblasts. ( A ) Overview of the sgRNA + 1 target site with sections of the sense and antisense (AS) ssODN that remove the c.828TAAA insertion and introduce a Kpn I recognition site by a silent C > A substitution. Predicted cut site for Cas9 are indicated by “ ˄ ” and “ ”, with “ ” indicating the strand that is nicked by Cas9D10A. Nucleotide numbers refer to the mutant c.828InsTAAA Fancf gene. ( B ) Overall gene editing efficiencies obtained in mouse fibroblasts depicted as general indel frequencies determined by TIDER (n = 3, one sided Student’s T-test, *p = 0.02). ( C ) Restriction fragment length polymorphism (RFLP) analysis by Kpn I digest of the Fancf locus provides evidence for template-based repair after Cas9, sgRNA + 1 and ssODN transfection (n = 3, one sided Student’s T-test, *p

    Journal: Scientific Reports

    Article Title: Effective CRISPR/Cas9-mediated correction of a Fanconi anemia defect by error-prone end joining or templated repair

    doi: 10.1038/s41598-018-36506-w

    Figure Lengend Snippet: CRISPR/Cas9 templated gene editing analysis in mouse fibroblasts. ( A ) Overview of the sgRNA + 1 target site with sections of the sense and antisense (AS) ssODN that remove the c.828TAAA insertion and introduce a Kpn I recognition site by a silent C > A substitution. Predicted cut site for Cas9 are indicated by “ ˄ ” and “ ”, with “ ” indicating the strand that is nicked by Cas9D10A. Nucleotide numbers refer to the mutant c.828InsTAAA Fancf gene. ( B ) Overall gene editing efficiencies obtained in mouse fibroblasts depicted as general indel frequencies determined by TIDER (n = 3, one sided Student’s T-test, *p = 0.02). ( C ) Restriction fragment length polymorphism (RFLP) analysis by Kpn I digest of the Fancf locus provides evidence for template-based repair after Cas9, sgRNA + 1 and ssODN transfection (n = 3, one sided Student’s T-test, *p

    Article Snippet: In experiments with a template ssODN for homology-directed repair Kpn I-HF (New England Biolabs) digestion reactions were performed on PCR product generated with the same primers.

    Techniques: CRISPR, Introduce, Mutagenesis, Transfection

    CRISPR/Cas9 templated gene editing analysis in mouse ES cells. ( A ) Overall gene editing efficiencies obtained in mouse embryonic stem cells depicted as general indel frequencies determined by TIDER (n = 3, one sided Student’s T-test, *p ≤ 0.02). ( B ) TIDE analysis of indel formation at predicted off-target loci after wtCas9 and sgRNA + 1 exposure irrespective of the ssODN orientation (n = 3, one sided Student’s T-test, *p ≤ 0.009). ( C ) Restriction fragment length polymorphism (RFLP) analysis by Kpn I digest of the Fancf locus provides evidence for template-based repair after Cas9 transfection (n = 6, one sided Student’s T-test, *p ≤ 0.02). ( D ) Mouse ES cells show enhanced clonal survival in the presence of 12.5 nM MMC after double-strand break formation by Cas9 and sgRNA + 1 independent of the orientation of the HDR template. A significant difference in relative mESC clone survival was also observed with the antisense (AS) ssODN following Cas9 nickase activity (n = 6, one sided Student’s T-test, *p ≤ 0.025), “M” = mock sgRNA.

    Journal: Scientific Reports

    Article Title: Effective CRISPR/Cas9-mediated correction of a Fanconi anemia defect by error-prone end joining or templated repair

    doi: 10.1038/s41598-018-36506-w

    Figure Lengend Snippet: CRISPR/Cas9 templated gene editing analysis in mouse ES cells. ( A ) Overall gene editing efficiencies obtained in mouse embryonic stem cells depicted as general indel frequencies determined by TIDER (n = 3, one sided Student’s T-test, *p ≤ 0.02). ( B ) TIDE analysis of indel formation at predicted off-target loci after wtCas9 and sgRNA + 1 exposure irrespective of the ssODN orientation (n = 3, one sided Student’s T-test, *p ≤ 0.009). ( C ) Restriction fragment length polymorphism (RFLP) analysis by Kpn I digest of the Fancf locus provides evidence for template-based repair after Cas9 transfection (n = 6, one sided Student’s T-test, *p ≤ 0.02). ( D ) Mouse ES cells show enhanced clonal survival in the presence of 12.5 nM MMC after double-strand break formation by Cas9 and sgRNA + 1 independent of the orientation of the HDR template. A significant difference in relative mESC clone survival was also observed with the antisense (AS) ssODN following Cas9 nickase activity (n = 6, one sided Student’s T-test, *p ≤ 0.025), “M” = mock sgRNA.

    Article Snippet: In experiments with a template ssODN for homology-directed repair Kpn I-HF (New England Biolabs) digestion reactions were performed on PCR product generated with the same primers.

    Techniques: CRISPR, Transfection, Activity Assay