	US 12,221,623 B2
	Methods of processing sperm for sex sorting
Thomas Boyd Gilligan, College Station, TX (US); Kenneth Michael Evans, College Station, TX (US); Richard Lenz, College Station, TX (US); Clara Gonzalez-Marin, Navasota, TX (US); and Ramakrishnan Vishwanath, Hamilton (NZ)
Assigned to INGURAN, LLC, Navasota, TX (US)
Filed by INGURAN, LLC, Navasota, TX (US)
Filed on Mar. 4, 2013, as Appl. No. 13/784,578.
Claims priority of provisional application 61/710,343, filed on Oct. 5, 2012.
Prior Publication US 2014/0099627 A1, Apr. 10, 2014
Int. Cl. C12N 5/076 (2010.01); C12N 5/071 (2010.01); C12Q 1/6806 (2018.01)

CPC C12N 5/061 (2013.01) [C12N 5/0612 (2013.01); C12Q 1/6806 (2013.01)]

22 Claims

1. A method comprising:

extending a bovine sperm sample in an initial extender at a sperm sample to initial extender ratio between about 1:1 and 1:10;

reconcentrating the extended bovine sperm sample to a concentration between 900 million sperm per ml and 2400 million sperm per ml, wherein the reconcentrated bovine sperm sample concentration is higher than the concentration of the extended bovine sperm sample following extension in the initial extender;

diluting the reconcentrated bovine sperm sample with a volume of staining buffer to a concentration between about 80 million sperm per ml and 320 million sperm per ml and adding a DNA selective dye;

staining the diluted bovine sperm sample;

determining the presence of an X or Y chromosome in the stained bovine sperm sample; and

collecting a population of viable X-chromosome bearing sperm and/or viable Y-chromosome bearing sperm.