GSM1194113: Gotheron Natural medium; Drosophila simulans; RNA-Seq
RNA extraction was performed using Macherey-Nagel Nucleospin RNA II kit, with 4 extraction per samples, that were then pooled. Quality and quantity were checked with an Experion (Biorad) RNA samples were precipitated and the pellets dissolved in water. The total RNAs were analysed for their integrity by capillary electrophoresis. From the total RNA samples poly(A)+ RNA was prepared which was used for cDNA synthesis. cDNA was synthesized using an oligo(dT)-linker primer and M-MLV H- reverse transcriptase for first strand synthesis. The reaction conditions were chosen such that the length of the first-strand cDNAs was limited to about 100 – 500 nt. Finally, the cDNAs were PCR amplified using a high fidelity DNA polymerase. For Solexa sequencing the cDNAs in the size range of 250 – 450 bp were eluted from preparative agarose gels. Aliquots of the size fractionated cDNAs were analyzed by capillary electrophoresis.
GEO Accession
GSM1194113