To generate mated females, females were paired with males until copulation occurred, at least 24 hr prior to pairing with virgin males.
To generate mated female targets in assays where males choose virgin or recently mated females, naive males were first paired with virgin females in 35 × 10 mm tissue culture dishes (Nunclon, Denmark) and observed for copulation.
Then we generated mate-pair reads with 1-2 Kb insert fragment size using Illumina HiSeq 2000 sequencing platform [ 26, 27] and used all mate-pair reads to join contigs into scaffolds.
In this work, we used microfluidics to generate mating pheromone gradients and tested the ability of yeast cells to sense and respond to the gradients by making a mating projection.
Our results agree with existing theory in that the opportunity for sexual selection is greater at higher male densities – females search through more males under those conditions, generating mating skew.
MHC allele-sharing (D, between the female and her social mate, and her genetic mate when she engaged in EP copulations) was compared with the distribution of expected D values obtained by randomly generating mating pairs (1000 bootstraps for each year, R version 2.15.0, R Development Core Team 2011).
The samples were then diluted and used for emulsion PCR. 10 ug of each DNA sample was used to generate mate-paired libraries with a 2.5 kb insert size using standard manufacturer protocols.
Of these 9,711 BESs, 4,441 BIBAC clones (84.3%) were successfully sequenced at both ends, generating mate-paired reads.
Of these 65,720 BES, 29,046 BAC clones (88.4%) were successfully sequenced on both ends, generating mate-pair reads.
A large-insert (~40 kb) fosmid library was constructed in the CopyControl™ pCCFOS™ vector system (Epicentre Technologies, USA).Corporation, USA) Insert ends (~800 bp/read) were sequenced generating mate pairs and 7.5 Mb sequencing data.
