Parus major transcriptome sequencing
Ten unrelated great tit fledglings were sampled from across the Wytham Woods population and brain, heart, kidney, liver, muscle, pancreas, skin and testes/ovaries dissected from each individual at between three and 23 minutes after death. Each chick was sexed at sampling and sex was later confirmed molecularly with the Z-002A marker. Tissues were stored in RNALater (Ambion) to prevent RNA degradation. RNA was extracted using the TRIzol® (Invitrogen) method. RNA quantity and degradation was tested using a Nanodrop (Agilent) with each tissue yielding a mean (SE) of 44.4 (7.5) ug of RNA with mean (SE) RNA integrity number (RIN) of 8.37 (0.21). For each tissue 1ug of RNA from each bird was pooled, i.e. to make 8 pools, each containing 10µg of RNA. Complementary DNA (cDNA) was synthesised by Evrogen (Moscow, Russia) from the eight tissue-specific pools using SMART kits. cDNA was normalised, also by Evrogen. cDNA sequencing was performed by the Liverpool Advanced Genomics Facility, with 3ug of normalised cDNA sequenced on a 454 FLX Titanium Genome Sequencer. Each tissue was sequenced on half a plate, i.e. four plates were sequenced in total.    tissue: number of sequences, total base pairs sequenced, mean sequence length, maximum sequence length  brain: 674719, 209572185, 311, 1796  heart: 408547, 112462495, 275, 1863  kidney: 655004, 198351347, 303, 1256  liver: 514017, 157069646, 306, 1005  muscle: 566729, 178678017, 315, 971  pancreas: 532017, 174131123, 327, 1726  skin: 573505, 154523708 269, 2028  testes: 663036, 199038872, 300, 1737  all tissues: 4587574, 1383827393, 302, 2028