Bombyx mori transcriptome sequencing project. Bombyx mori

Total RNAs were prepared using Trizol reagent (Invitrogen) according to the manufacturer's protocol. Total RNA (10 ug) was loaded onto a 15% denaturing polyacrylamide gel containing 10 M urea, electrophoresed, and then stained with SYBRGold (Invitrogen). Signals were visualized using LAS-1000 film (Fujifilm). Small RNA libraries were constructed using a Small RNA cloning kit (Takara). DNA sequencing was performed using the Solexa genetic analysis system (Illumina) (Bently, 2006). One nano-gram of the prepared cDNA was used for the sequencing reactions with the Illumina GA. 10,000-15,000 clusters were generated per 'tile' and 36 cycles of the sequencing reactions were performed. The protocols of the cluster generation and sequence reactions were according to the manufacturer's instructions. Solexa sequencing generated reads of up to 36 nucleotides in length. cell line/type' 1:Wild type strain p50T, day 4 pupal ovary (called 'OV' in this study) 2:Wild type strain p50T, day 4 pupal testis (called 'TE' in this study) 3:Sex-limited yellow(T(W:2)Y-Abe; called 'LY' in this study) , day 4 pupal ovary 4:Mandarina W, (called 'MW' in this study), day 4 pupal ovary 5:DfZ-DfW (called 'without Fem', WF in this study), day 4 pupal testis