GSM2254810: PTTH 3 hr; Bombyx mori; RNA-Seq
Total RNA was extracted by using the TRIzol Reagent (Invitrogen) and RNeasy Mini Kit (Qiagen).  The frozen PGs were homogenized in 500 microliter of TRIzol Reagent and incubated at room temperature for five minutes before adding 350 microliter of buffer RLT (QIAGEN) containing 1% beta-mercaptoethanol.  Samples were vortexed and 100 microliter of chlorophorm was added before spinning at 12,000 rpm for 15 min at 4ºC.  The aqueous phase was removed and RNA was isolated according to the instruction of RNeasy Mini Kit. RNA sequencing libraries were prepared according to specifications by the University of Minnesota’s Biomedical Genomics Center (BMGC) using the Paired-end mRNA-Seq 8-Sample Prep Kit (Illumina).