GSM8194353: Brain, Paxton limnetic, conspecific courtship, trial 20; Gasterosteus aculeatus; RNA-Seq
After 20 min of time with the courting male/ female social control or spawning (whichever came first), the female subject was killed by rapid cervical dissection, and the brain was immediately dissected under a microscope in a Sylgaard 184-lined petri-dish filled with a Ringer's solution. Brains were stored overnight in RNAlater at 4°C and then transferred to -20°C until RNA extraction. Brains were shipped on dry ice from Michigan State University to The University of Texas at Austin where RNA was extracted using the Maxwell 16 LEV simplyRNA Tissue Kit, which utilizes a robot to increase consistency. At the UT Austin Genomic Sequencing and Analysis Facility (GSAF) each sample (all with RIN > 7.8, 8.8 ± 0.4 mean ± SD, 11 samples do not have RIN scores) was prepared for RNA-seq using Poly­A mRNA capture and given a unique barcode. A single library of all multiplexed samples was sequenced across eight lanes of an Illumina Hiseq 2500 with 2x50 PE chemistry.