2.5 | Chromosome preparation and in situhybridization
Chromosomes were prepared for GISH analysis according to the method of
Han et al. (2004). Total genomic DNA was extracted from fresh leaves by
the CTAB method (Murray & Thompson, 1980). Plasmids (from positive
clones that are St genome) and the StY genome were
labeled with fluorescein-12-dUTP or Texas-red-5-dCTP using the nick
translation method. Hybridization procedure, detection, and
visualization were performed according to the method of Wang et al
(2017).