GSM1603396: 3_8; Drosophila melanogaster; RNA-Seq
Ovarian follicles of the indicated stage and genotype were dissected under a dissecting microscope at room temperature in Grace's medium and transferred to iced tripure (Roche/Boehringer Mannheim cat: 11667157001) to avoid RNA degradation. Samples of 100 follicles were homogenized. 600μl of fresh tripure was added, mixed, and allowed to stand at room temperature for 5-10 min. After adding 180μl chloroform, samples were vortexed 2 x 45 secs, and incubated at room temperature for 10 minutes. Samples were then centrifuged for 15 min at 12,000 rpm at 4°C, and the aqueous layer was moved to a fresh tube. RNA was precipitated by adding 400μl of isopropanol, vortexing for 15 seconds, and incubating at room temperature for 15 min, and centrifuging at 12,000 rpm for 15 min at 4°C. After washing the pellet with 75% EtOH, the RNA was dried in air dry for 5 minutes, resuspended in 50μl nuclease free H2O, and stored at -80°C. cDNA libraries were constructed from poly(A)-selected RNA using Illumina TruSeq RNA Library Prep Kit v2
GEO Accession
GSM1603396