GSM1482518: Df(1)ED6630_F_Df(1)/w1118_rep_3 [Sample_101103]; Drosophila melanogaster; RNA-Seq
We used RNeasy 96 Kit (Qiagen, Valencia, CA) to extract total RNA from a pool of 10 dissected heads for each sample. Frozen heads were added to 2ml Axygen 96-well plates (Corning, Life Sciences, Union City, CA) pre-loaded with 200 µL 1mm glass beads (Biospec Products, Inc., Bartlesville, OK) and 200 µL RLT Buffer (Qiagen, Valencia, CA). After covering the plates with Axygen sealing mats, we homogenized the samples with Mini-BeadBeater-96 (Biospec Products Inc., Bartlesville, OK). Manual purification was carried out using QIAvac 96 vacuum manifold (Qiagen, Valencia, CA). We followed TruSeq RNA Sample Preparation V2 high-throughput protocol (Illumina Inc., San Diego, CA) to construct RNA-seq libraries from polyA+ RNA. For each sample, 100ng of total RNA was used as input in the initial step and 10pg ERCC spike-in RNA (Zook et al. Plos ONE 2012, 7(7): e41356., pool 78A) was added to the Elute, Prime, Fragment Mix in the "Purify and fragment mRNA” step.
GEO Accession
GSM1482518