	US 12,215,309 B2
	Identification of rare products of crossing organisms
Arthur Roelof Gorter De Vries, Delft (NL); Charlotte Catharina Koster, Delft (NL); Jean-Marc Georges Daran, Delft (NL); Jan-Maarten Geertman, Amsterdam (NL); and Niels Gerard Adriaan Kuijpers, Amsterdam (NL)
Assigned to Heineken Supply Chain B.V., Amsterdam (NL)
Appl. No. 17/054,271
Filed by Heineken Supply Chain B.V., Amsterdam (NL)
PCT Filed May 10, 2019, PCT No. PCT/NL2019/050279 § 371(c)(1), (2) Date Nov. 10, 2020, PCT Pub. No. WO2019/216769, PCT Pub. Date Nov. 14, 2019.
Claims priority of application No. 2020912 (NL), filed on May 11, 2018.
Prior Publication US 2021/0246415 A1, Aug. 12, 2021
Int. Cl. C12N 1/16 (2006.01); C12Q 1/6895 (2018.01); C12R 1/865 (2006.01); G01N 1/30 (2006.01)

CPC C12N 1/16 (2013.01) [C12Q 1/6895 (2013.01); G01N 1/30 (2013.01); C12R 2001/865 (2021.05)]

10 Claims

1. A method for identifying a hybrid yeast organism, comprising

(a) providing cells from a first and second yeast parent organism,

whereby the first and second yeast parent organisms are mating-compatible,

whereby the first yeast parent organism differs from the second yeast parent organism,

whereby said first yeast parent organism, but not said second yeast parent organism, carries an auxotrophic marker,

(b) labelling cells from the second yeast parent organism with a fluorescent dye;

(c) hybridizing cells from the first yeast parent organism with labelled cells from the second yeast parent organism at a temperature between 5° C. and 15° C.; and

(d) identifying a hybrid yeast organism as an auxotrophic, labelled cell.