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
First Claim
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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Abstract
A method for determining the integrity of the genome of a sample and/or the quality of a library of DNA sequences obtained by deterministic restriction site whole genome amplification can include (a) amplifying the library of DNA sequences to produce first, second, and third PCR products each of a different size from 50 bp to 1000 bp, by PCR using at least one first primer pair, one second primer pair and one third primer pair, the primer pairs each hybridizing to a DNA sequence of the library having a length from 1000 bp to 5000 bp and corresponding to a sequence of the genome located respectively on a first, second and third chromosome arm; (b) detecting the first, second and third PCR products; (c) correlating the presence of the first, second and third PCR products with the integrity of the genome of the sample and/or the quality of the library.
1 Citation
10 Claims
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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:
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(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, and at 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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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Specification