Whole transcriptome profiling of the rat pineal gland DCK2
For the mRNA-Seq sample preparation, the Illumina standard kit was used according to the manufacturer's protocol. Briefly, 10 microgram of each total RNA sample was used for polyA mRNA selection using streptavidin-coated magnetic beads, followed by chemical mRNA fragmentation. The fragmented mRNA was subjected to cDNA synthesis using reverse transcriptase (SuperScript II) and random primers.  The cDNA was further converted into double stranded cDNA and, after an end repair process (Klenow fragment, T4 polynucleotide kinase and T4 polymerase), was finally ligated to Illumina paired end (PE) adaptors. Size selection was performed using a 2% agarose gel. Libraries were enriched using 15 cycles of PCR and purified by the QIAquick PCR purification kit (Qiagen). The enriched libraries were diluted with Elution Buffer to a final concentration of 10 nM. Each library was run at a concentration of 7 pM on one Illumina Genome Analyzer II (GAii) lane using paired-end 51 base-pair sequencing. Image analysis and base calling was done by Illumina Genome Analyzer Pipeline software (GAPipeline) with default parameters.