title: GSM1186463: 3 day old heads normal food 2; Drosophila melanogaster; RNA-Seq
library_construction_protocol: Spd- and Spd+ flies were frozen in liquid nitrogen and heads were collected after vortexing by mechanical separation. Poly A+ RNA was extracted from approximately 50 ul of heads per sample using the RNeasy Lipid tissue Mini kit (Qiagen) and the Oligotex mRNA mini kit (Quiagen). RNA was reverse transcribed into a cDNA using the Superscript III Reverse Transcriptase Kit (Invitrogen) for first strand synthesis. Second strand synthesis was performed with E.coli and T4 DNA polymerases (New England Biolabs). DNA end repair was performed on 9-20 ng of cDNA followed by ligation of adaptors, and size selection for 300 bp fragments using the E-gel Size Select gel (Invitrogen). Fragments were amplified using 14 PCR cycles and linear amplification was validated by qPCR.
attributes.tag: GEO Accession
attributes.value: GSM1186463