	US 12,442,031 B1
	DNA labeling compositions and methods for comparative genomic hybridization
Praveen Pande, Holbrook, NY (US); Jack Coleman, East Northport, NY (US); and Christian Merz, Hicksville, NY (US)
Assigned to Enzo Biochem, Inc., Farmingdale, NY (US)
Filed by Enzo Biochem, Inc., New York, NY (US)
Filed on Jun. 18, 2018, as Appl. No. 16/010,999.
Claims priority of provisional application 62/510,997, filed on May 25, 2017.
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6806 (2018.01); C12Q 1/6813 (2018.01); C12Q 1/6876 (2018.01); C40B 50/04 (2006.01)

CPC C12Q 1/6806 (2013.01) [C12Q 1/6813 (2013.01); C12Q 1/6876 (2013.01); C12Q 2523/10 (2013.01); C12Q 2533/101 (2013.01); C12Q 2563/107 (2013.01); C40B 50/04 (2013.01)]

12 Claims

1. A method for preparing fluorescently labeled output DNA suitable for array-based comparative genomic hybridization from input genomic DNA, comprising the steps of:

providing a sample of genomic DNA in an amount of 50+/−25% nanograms;

admixing the sample of genomic DNA with random DNA primers in a reaction buffer comprising magnesium,

wherein the admixture does not contain either of ethylenediaminetetraacetic acid (EDTA) and egtazic acid (EGTA);

heating the admixture to denature the genomic DNA and then cooling the admixture to permit the random DNA primers to anneal to the genomic DNA in the admixture;

further admixing to form a primer extension composition comprising

a Klenow fragment DNA-dependent DNA polymerase,

0.4 mM+20% dATP,

0.4 mM+20% dCTP,

0.4 mM+20% dGTP,

0.4 mM+20% dTTP,

0.08 mM+20% to 0.16 mM+20% fluorescently labeled dUTP,

wherein the primer extension composition formed does not contain either of ethylenediaminetetraacetic acid (EDTA) and egtazic acid (EGTA); and

incubating the primer extension composition at a temperature permissive for primer extension by the polymerase to extend the random DNA primers on the genomic DNA, thereby forming fluorescently labeled DNA extension products.