GSM1603251: SNS-Seq (pool) from S2 cells, pooling replicates 1 and 2; Drosophila melanogaster; OTHER
SNS fragments were isolated essentially as described in (Cayrou et al. 2011). Adjustments were made according to SW-41 Ti rotor specifications (Beckman-Coulter) and centrifugation was carried out at 4 °C and 26700 rpm for 21 hours. DNA fragments of 0.5-2.5 kb were collected, purified and subjected to up to five rounds of T4 PNK (Fermentas) phosphorylation and lambda Exonuclease (Fermentas) digestion. SNS were then prepared for sequencing by digesting the RNA-primer, second strand synthesis (NEBNext mRNA Second Strand Synthesis) and purified with AMPure XP beads (Beckman-Coulter). DNA libraries were prepared with the Nextera XT DNA Sample Preparation Kit (Illumina) following manufacturer’s instructions.
GEO Accession
GSM1603251