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Method and kit for determining the genome integrity and/or the quality of a library of DNA sequences obtained by deterministic restriction site whole genome amplification

  • US 10,392,657 B2
  • Filed: 12/04/2014
  • Issued: 08/27/2019
  • Est. Priority Date: 12/04/2013
  • Status: Active Grant
First Claim
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1. A method for determining the integrity of the genome of a sample to be analysed by whole genome amplification (WGA) and/or the quality of a library of DNA sequences obtained by deterministic restriction site whole genome amplification (DRS-WGA) of the genome of the sample comprising the steps of:

  • (a) providing a sample having a genome(b) obtaining a library of DNA sequences by deterministic restriction site whole genome amplification (DRS-WGA) of the genome of the sample;

    (c) amplifying the library of DNA sequences by PCR using;

    at least one first primer pair which hybridizes to a DNA sequence of the library having a length from 1000 bp to 5000 bp and encompassing the D5S2117 region of chromosome 5q,at least one second primer pair which hybridizes to a DNA sequence of the library having a length from 1000 bp to 5000 bp and encompassing exons 2 and 3 of the TRP53 gene of the genome, andat least one third primer pair which hybridizes to a DNA sequence of the library having a length from 1000 bp to 5000 bp and encompassing the KRT19 pseudo-gene 1 of the genome,wherein the step of amplifying giving rise to a first PCR product from 50 bp to 1000 bp, a second PCR product from 50 bp to 1000 bp having a size other than the first PCR product, and a third PCR product from 50 bp to 1000 bp having a size other than the first and second PCR products; and

    (d) detecting the first, second and third PCR products,wherein the presence of one or more of the first, second and third PCR products corresponds to a level of the integrity of the genome of the sample and/or the quality of the library of DNA sequences.

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