GSM7814629: head, old, female, H3.3Adelta, rep2; Drosophila melanogaster; RNA-Seq
Heads were removed from the rest of the carcass and total RNA was extracted. Briefly, 10-25 flies were anesthetized and placed in a 15 mL tube. The tube was submerged in liquid nitrogen for 5-10 seconds and then vortexed for 5-10 seconds to separate heads from carcasses. This process was repeated for a total of 5 times before pouring the contents of the tube into a set of stacked sieves. Fly heads were crushed in TRIzol reagent (Thermo Fisher) and RNA was purified with the RNA clean & concentrator-5 kit (Zymo research). Total (Ribo-minus) RNA-seq libraries, three replicates per genotype, were prepared using a Universal RNA-seq library preparation kit with NuQuant (Tecan). Sequencing was performed on a NovaSeq 6000SP (Illumina) producing 100 nt, paired-end reads.