US 9,809,794 B2
Materials and methods for achieving differential lysis of mixtures with the aid of alkaline lysis and pressure cycling technology (PCT)
Bruce R. McCord, Miami, FL (US); Deepthi V. Nori, Miami, FL (US); Nathan P. Lawrence, South Easton, MA (US); and Richard Schumacher, South Easton, MA (US)
Assigned to THE FLORIDA INTERNATIONAL UNIVERSITY BOARD OF TRUSTEES, Miami, FL (US); and PRESSURE BIOSCIENCES, INC., South Easton, MA (US)
Filed by The Florida International University Board of Trustees, Miami, FL (US)
Filed on Feb. 18, 2015, as Appl. No. 14/625,233.
Claims priority of provisional application 61/941,201, filed on Feb. 18, 2014.
Prior Publication US 2015/0232917 A1, Aug. 20, 2015
Int. Cl. C12Q 1/68 (2006.01); C12N 1/06 (2006.01); C12N 15/10 (2006.01)
CPC C12N 1/06 (2013.01) [C12N 15/1003 (2013.01)] 14 Claims
 
1. A method of separating male sperm cells and female epithelial cells in a sample embedded in a cotton swab, comprising:
immersing the sample in an alkaline solution comprising 0.2-0.8 N NaOH,
wherein said alkaline solution does not comprise deoxyribonuclease (DNase);
applying pressure cycling technology (PCT) to the sample at room temperature, the PCT comprising the application of alternating cycles of ambient and high pressure to the sample, said high pressure being between 10,000 psi and 45,000 psi;
neutralizing the sample with a buffer solution;
centrifuging the sample to produce an eluted epithelial fraction;
collecting and purifying the eluted epithelial fraction for female DNA processing, whereby a sperm fraction is left in the cotton swab; and
processing the sperm fraction remaining in the cotton swab.