Brains were removed, placed in RNAlater overnight at 4C. Excess RNAlater was subsequently removed and the brains stoed at -80C until total RNA extraction. Total RNA was extracted using RNeasy Plus Mini kit (Qiagen). We pooled 1 ug of total RNA from 10 same sex and same strain individuals into one biological replicate. We followed Illumina TruSeq RNA Sample Prep V2 protocol using 1 ug of total RNA for the construction of sequencing libraries.
RNA libraries were prepared for sequencing using standard Illumina protocols for multiplexing. Adapter sequences are as follows: lsbf1 (GTGAAA(C)); lsbf2 (GTTTCG(G)); lsbm1 (GTCCGC(A)); lsbm2 (GTGGCC(T)); hsbf1 (CCGTCC(C)); hsbf2 (GGCTAC(A)); hsbm1 (ATGTCA(G)); hsbm2 (TAGCTT(A)); abf1 (ATTCCT(T)); abf2 (GATCAG(A)); abm1 (ACTGAT(A)); abm2 (ACTTGA(A)); shf1 (GAGTGG(A)); shf2 (TTAGGC(A)); shm1 (CGTACG(T)); shm2 (ATCACG(A))