title: GSM2935156: Oncopeltus_fasciatus_dsDNMT1_RNA-seq_biological_rep_3_of_3; Oncopeltus fasciatus; RNA-Seq
library_construction_protocol: Ovaries were removed from each female, flash frozen in liquid nitrogen and stored at -80°C until processing. Total RNA and genomic DNA was extracted from pairs of ovaries (from a single female) using a Qiagen RNAeasy micro kit (Qiagen, Venlo, The Netherlands) per manufacturer's instructions with Qiagen Qiazol as the lysis buffer. Complementary DNA (cDNA) was synthesized from 500 ng RNA with qScript cDNA SuperMix (Quanta Biosciences, Gaithersburg, MD). DNA methylation. MethylC-seq libraries were prepared using the protocol described in Urich et al. (2015; doi:10.1038/nprot.2014.114). Lambda phage DNA was used as a control for the sodium bisulfite non-conversion rate. Gene expression. RNA-seq libraries for RNA extracted from three DNMT1 knocked down and three control individuals were constructed using Illumina TruSeq Stranded RNA LT Kit (Illumina, San Diego, CA) following the manufacturer's instructions with limited modifications. The starting quantity of total RNA was adjusted to 1.3 µg, and all volumes were reduced to a third of the described quantity.
attributes.tag: GEO Accession
attributes.value: GSM2935156