alias: GSM2318692
title: GSM2318692: 36G; Apis mellifera; RNA-Seq
library_construction_protocol: The groups included in the gene expression study included well distinguished bees of each the three behavioural types: nurses, guards, and unresponsive bees. The bees were collected by flash freezing in liquid nitrogen and were placed in marked 1.5 mL microcentrifuge tube in a dry ice bucket. The bees were kept at -80° C until tissue collection. Six groups of bees from each of two SDI colonies were included in the gene expression analysis. Bees’ heads were separated from bodies on dry ice and placed in a dissection dish with 200 proof ethanol and dry ice to prevent RNA degradation during dissection. The cuticle of the head capsule was removed and each head was placed in RNAlaterICE (Life Technologies) at -20ºC for 14 h to 18 h. The heads were then opened, the hypopharyngeal glands removed, and the whole brain removed. Using a fine scalpel, the optic lobes were removed and a horizontal incision was made across the midbrain through the posterior protocerebral lobe. The lower part of the midbrain containing the antennal lobes and the subesophageal ganglion were removed. The upper part of the midbrain containing mostly the mushroom bodies (MB) and the central brain was used for gene expression analysis. MB were placed in a new 1.5 mL microcentrifuge tube and kept frozen at -80° C until RNA extraction. RNA extraction was performed with a PicoPure RNA Isolation Kit (Applied Biosystems; Lot #1210063) according to the manufacturer’s specifications, including a DNase treatment (QIAGEN) to remove genomic DNA contamination. 550 ng RNA from each sample were used for whole mRNA expression analysis. The RNA-Seq libraries were constructed with the TruSeq® Stranded mRNA HT (Illumina) using an ePMotion 5075 robot (Eppendorf) according to manufacturer's specifications. The libraries were pooled in equimolar concentration as instructed and each pool was quantitated by qPCR and sequenced  for 101 cycles on a HiSeq2500 using a HiSeq SBS sequencing kit v. 4.
identifiers.namespace: GEO
identifiers.id: GSM2318692