| US 7,351,529 B2 | ||
| Methods for detecting genetic mosaicisms using arrays | ||
| Mansoor Mohammed, Houston, Tex. (US) | ||
| Assigned to PerkinElmer LAS, Inc., Waltham, Mass. (US) | ||
| Filed on Sep. 27, 2002, as Appl. No. 10/260,733. | ||
| Claims priority of provisional application 60/325853, filed on Sep. 27, 2001. | ||
| Prior Publication US 2003/0124584 A1, Jul. 03, 2003 | ||
| Int. Cl. C12Q 1/68 (2006.01); C12P 19/34 (2006.01); C07H 21/04 (2006.01) | ||
| U.S. Cl. 435—6 [435/91.2; 536/24.31; 536/24.32; 536/24.33] | 40 Claims |
| 1. A method of detecting a degree of genetic mosaicism in a cell population by performing an array-based comparative genomic
hybridization (CGH), wherein an array comprising a plurality of cloned genomic nucleic acid segments is provided in a plurality
of identical replicas, each cloned segment immobilized to a discrete and known spot on a substrate surface to form the array,
the cloned genomic nucleic acid segments comprising a substantially complete first genome of a known first karyotype, the
method comprising:
(a) contacting replicas of the array with mixtures of a first nucleic acid sample and a second nucleic acid sample and fractional
dilutions of the second sample, wherein the first sample comprises a plurality of genomic nucleic acid segments comprising
a substantially complete complement of the first genome labeled with a first detectable label, the second sample-comprises
a plurality of genomic nucleic acid segments comprising a substantially complete complement of the second genome labeled with
a second detectable label, and the karyotype of the second sample is known and is different from that of the first sample;
(b) contacting further replicas of the array with mixtures of the first nucleic acid sample and a third nucleic acid sample
and fractional dilutions of the third sample, wherein the third sample comprises a genomic nucleic acid sample with an unknown
karyotype and is labeled with the second detectable label, and the genomic nucleic acid of the third sample comprises a substantially
complete complement of genomic nucleic acid of a third genome from a test cell or a tissue sample, wherein the contacting
is under conditions wherein the nucleic acid in the mixtures of each of the first and second samples and the first and third
samples can specifically hybridize to the genomic nucleic acid segments immobilized on the array;
(c) measuring the amount of first label and second label on each spot for each respective contacted array and determining
the karyotype of each dilution fraction by comparative genomic hybridization; and,
(d) selecting which fractional dilution karyotype determination of the second sample most closely determines the known karyotype,
and selecting data for the same fractional dilution of the third sample to determine the karyotype of the third sample, thereby
determining the degree of genetic mosaicism in the-cell population.
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