dna substrates φx174 single stranded ss virion dna  (New England Biolabs)


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    Structured Review

    New England Biolabs dna substrates φx174 single stranded ss virion dna
    HsRAD51(F129V) and HsRAD51(H294V) are deficient in D-loop formation and strand exchange. ( A ) In vitro D-loop assay reaction schematic. ( B ) 0.8 µM of HsRAD51, HsRAD51(F129V), or HsRAD51(H294V) and [P 32 ]-labeled ssDNA (90mer; 2.4 µM nt) were preincubated for 10 min at 37°C in the reaction buffer containing 1 mM ATP and 1 mM MgCl 2 or CaCl 2 . Reactions were initiated by the addition of supercoiled pBS SK(-) plasmid <t>DNA</t> (35 µM bp). After 15 min, reactions were terminated by the addition of proteinase-K and SDS. Joint molecules (JMs) were analyzed on a 0.9% agarose gel. ( C ) Analysis of salt and RPA requirement for strand exchange. Reaction schematic shown above: HsRAD51 (5 µM) and <t>φX174</t> circular ssDNA (30 µM nt) were pre-incubated with 2.5 mM ATP and 1 mM MgCl 2 at 37°C for 5 m prior to the addition of 150 mM NaNH 4 HPO 4 (if indicated) and linear φX174 dsDNA (15 µM bp). After 5 m, HsRPA (2 µM) was added (if indicated) and the incubation was continued for 3 h. Samples were deproteinized and analyzed on 0.9% agarose gel with 0.1 µg/mL ethidium bromide.
    Dna Substrates φx174 Single Stranded Ss Virion Dna, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna substrates φx174 single stranded ss virion dna/product/New England Biolabs
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    dna substrates φx174 single stranded ss virion dna - by Bioz Stars, 2020-12
    85/100 stars

    Images

    1) Product Images from "Subunit Interface Residues F129 and H294 of Human RAD51 Are Essential for Recombinase Function"

    Article Title: Subunit Interface Residues F129 and H294 of Human RAD51 Are Essential for Recombinase Function

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0023071

    HsRAD51(F129V) and HsRAD51(H294V) are deficient in D-loop formation and strand exchange. ( A ) In vitro D-loop assay reaction schematic. ( B ) 0.8 µM of HsRAD51, HsRAD51(F129V), or HsRAD51(H294V) and [P 32 ]-labeled ssDNA (90mer; 2.4 µM nt) were preincubated for 10 min at 37°C in the reaction buffer containing 1 mM ATP and 1 mM MgCl 2 or CaCl 2 . Reactions were initiated by the addition of supercoiled pBS SK(-) plasmid DNA (35 µM bp). After 15 min, reactions were terminated by the addition of proteinase-K and SDS. Joint molecules (JMs) were analyzed on a 0.9% agarose gel. ( C ) Analysis of salt and RPA requirement for strand exchange. Reaction schematic shown above: HsRAD51 (5 µM) and φX174 circular ssDNA (30 µM nt) were pre-incubated with 2.5 mM ATP and 1 mM MgCl 2 at 37°C for 5 m prior to the addition of 150 mM NaNH 4 HPO 4 (if indicated) and linear φX174 dsDNA (15 µM bp). After 5 m, HsRPA (2 µM) was added (if indicated) and the incubation was continued for 3 h. Samples were deproteinized and analyzed on 0.9% agarose gel with 0.1 µg/mL ethidium bromide.
    Figure Legend Snippet: HsRAD51(F129V) and HsRAD51(H294V) are deficient in D-loop formation and strand exchange. ( A ) In vitro D-loop assay reaction schematic. ( B ) 0.8 µM of HsRAD51, HsRAD51(F129V), or HsRAD51(H294V) and [P 32 ]-labeled ssDNA (90mer; 2.4 µM nt) were preincubated for 10 min at 37°C in the reaction buffer containing 1 mM ATP and 1 mM MgCl 2 or CaCl 2 . Reactions were initiated by the addition of supercoiled pBS SK(-) plasmid DNA (35 µM bp). After 15 min, reactions were terminated by the addition of proteinase-K and SDS. Joint molecules (JMs) were analyzed on a 0.9% agarose gel. ( C ) Analysis of salt and RPA requirement for strand exchange. Reaction schematic shown above: HsRAD51 (5 µM) and φX174 circular ssDNA (30 µM nt) were pre-incubated with 2.5 mM ATP and 1 mM MgCl 2 at 37°C for 5 m prior to the addition of 150 mM NaNH 4 HPO 4 (if indicated) and linear φX174 dsDNA (15 µM bp). After 5 m, HsRPA (2 µM) was added (if indicated) and the incubation was continued for 3 h. Samples were deproteinized and analyzed on 0.9% agarose gel with 0.1 µg/mL ethidium bromide.

    Techniques Used: In Vitro, Labeling, Plasmid Preparation, Agarose Gel Electrophoresis, Recombinase Polymerase Amplification, Incubation

    Related Articles

    Gel Extraction:

    Article Title: Subunit Interface Residues F129 and H294 of Human RAD51 Are Essential for Recombinase Function
    Article Snippet: .. DNA Substrates φX174 single-stranded (ss) virion DNA and replicative form I (RFI) were purchased from NEB. φX174 RFIII was obtained by linearizing RFI with ApaLI restriction enzyme and gel purifying with Qiaquick Gel Extraction kit (Qiagen). .. For surface plasmon resonance (SPR) analysis, a 5′ biotinylated oligo dT50 was used as ssDNA and for dsDNA 5′ biotinylated 50-mer 5′-TCG AGA GGG TAA ACC ACA- ATT ATT GAT ATA AAA TAG TTT TGG GTA GGC GA was annealed with its complement.

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    New England Biolabs neb φx174 rfiii
    Neb φx174 Rfiii, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/neb φx174 rfiii/product/New England Biolabs
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    neb φx174 rfiii - by Bioz Stars, 2020-12
    85/100 stars
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