title: GSM4827611: Amel_embryo_25-40h_female_rep2; Apis mellifera; RNA-Seq
library_construction_protocol: Total RNA was isolated from pools of 30 (55-70 h old embryos), 60 (25-40 h old embryos) or 100 (10-15 h old embryos) from three different mothers . 250 µl Trizol were added to each pool and the tissue was homegenized using a pestle. After incubation for 5 min at room temperature 50 µl chloroform were added. The solution was mixed for 30 sec and centrifuged for 15 min on 13,000 rpm at 4°C. The upper phase was transferred into a new tube and 100 µl 70% ethanol were added, mixed and transfered to a purification column (Qiagen RNeasy Mini Kit, Qiagen). The purification was performed accoring to protocol with an included on-column DNaseI treatment (10 µl DNase and 70 µl buffer RDD; RNAse-Free DNase Set, Qiagen) for 15 minutes.