We sought to investigate whether the Rosetteless phenotype was dominant or recessive by examining the phenotypes of the heterozygotes isolated after mating Rosetteless to Isolate B. All seven heterozygous cultures contained predominantly thecate cells a cell type that is not competent for rosette formation (Dayel et al., 2011)—and three of these cultures also contained a small number of rosettes.
For larger-scale analyses, however, efficiency would be significantly enhanced by the use of selectable markers under the control of the mating-pheromone promoters [ 61] to isolate mating-competent diploids, and another marker to select tetraploids [ 27].
Twelve remaining fzo1 ∆ aac2 ∆ cyh2 sfa isolates were mated to cyh2 strain CDD51 transformed with plasmid b19 and lacking functional mtDNA following two days of culture in the presence of 25 µg/mL ethidium bromide (Goldring et al. 1970).
The eventual second-placed rider, Pierre Rolland, stalled briefly, but Bardet's view was "there were two Europcar riders" – Rolland and his team-mate Cyril Gautier – "I had to isolate them.
To isolate zygotes, mice were mated with C57Bl/6 males and checked for the presence of a copulatory plug on the following morning.
Each line was starved, mated to each of the others and 44 single mating pairs isolated into separate drops of standard growth medium (Hamilton and Orias, 2000).
However, if isolation from mushroom tissues still fails, there might still be the possibility for heterothallic species to generate a new dikaryotic mycelium from poly-spore germination on agar plates or by mating of isolated germinated basidiospores (Schuytema et al. 1966; Snelling et al. 1996; Lodge et al. 2004).
L4 unc-31 hermaphrodites were staged and used 3.5 days later for mating with isolated 18 24-hour 18 24-hourrgin adult males.
Such differential viability may also explain the low rate of outcrossed mating, if few Rosetteless haploids survived to mate with Isolate B haploids.
After mating, females were isolated in individually marked cages surrounding live tree branches suitable for oviposition and feeding.
