US 11,781,127 B2
Homogeneous method to prepare sperm DNA from sexual assault cases
Alex Garvin, Durmenach (FR)
Filed by Alex Garvin, Durmenach (FR)
Filed on Jan. 11, 2008, as Appl. No. 12/8,385.
Claims priority of provisional application 60/880,497, filed on Jan. 16, 2007.
Prior Publication US 2008/0261293 A1, Oct. 23, 2008
Int. Cl. C12N 15/10 (2006.01)
CPC C12N 15/1003 (2013.01) 6 Claims
 
1. A method of separating human sperm DNA from human non-sperm DNA in sexual assault samples comprising:
contacting a sample containing human sperm cells and human non-sperm cells with proteinase-K in an extraction buffer solution comprising t-octylphenoxypolyethoxyethanol, wherein the extraction buffer solution does not contain sodium dodecyl sulfate;
digesting the human non-sperm cells with the proteinase-K;
adding a DNAse I to the same extraction buffer solution in which the non-sperm cells were digested without a centrifugation step or a filtration step with respect to the addition of the DNAse I;
selectively degrading non-sperm DNA from the non-sperm cells with the DNAse I;
incubating the DNAse I and lysing the sperm by adding ethylenediaminetetraacetic (EDTA) and dithiothreitol (DTT) at 56° C. until the DNAse I is inactivated; and
separating sperm DNA from the human sperm for further analysis of the sperm DNA.