Umbrella record for two collections (male and female) of short sequencing reads of cDNA prepared from polyadenylated RNA isolated from heads of Drosophila pseudoobscura adults. Transcriptome represented as frequency of reads along genome (see GBrowse presentation; data in wiggle format); restricted to uniquely aligned reads.
Adults were aged 8 days post-eclosion and sorted by sex. Collection of adult heads: frozen whole flies were placed on 10.8 x 10.8 cm ceramic plates frozen on dry ice; forceps were used to dissect 400-500 heads for subsequent RNA extraction.
mRNA was purified from total RNA using a QIAGEN Oligotex mRNA kit (Valencia, California). mRNA quality and quantity was determined by Bioanalyzer (Agilent, Santa Clara, California) and 260/280 ratio from Nanodrop ND-1000 UV spectrophotometer. Double-stranded cDNAs were made from mRNA using the Superscript II (Invitrogen) and random hexamer primers (Invitrogen). DNA fragment libraries were prepared for sequencing by addition adaptor oligonucleotides; cDNA templates were size-selected by gel purification (200bp +/- 25bp), then amplified by PCR using oligo-specific primers and Phusion polymerase. The amplified product was purified using a QIAquick column.
Read length (bases):75
Only Illumina GAII data was used. Raw reads were trimmed to 75 nt before mapping (shorter reads were removed) and mapped using TopHat (v2.0.8b) with parameters: -g 1 -r 150 --solexa1.3-quals (except for data from head samples where parameters were -g 1 -r 150).
Strain used: line MV2-25 Baylor sequencing strain, obtained from the Drosophila Species Stock Center (DSSC).
[Transcriptional Profiling of additional Drosophila species](http://intermine.modencode.org/release-33/experiment.do?experiment=Transcriptional+Profiling+of+additional+Drosophila+species+with+RNA-Seq)