title: GSM2975536: S2 cell (ExpressFive medium) smallRNA-seq replicate 2; Drosophila melanogaster; ncRNA-Seq
library_construction_protocol: SmallRNA libraries from samples were generated using NEXTflex Small RNA Library Prep kit v3 (Bioo Scientific #5132), starting with 2 µg of total RNA and SRQC/ERDN-spike-in mix (Locati et al. 2015) (a generous gift from Dr. Timo Breit's lab at University of Amsterdam) using the following modifications: Following upon excessive 3' adapter removal the samples were eluted in 12 µl H2O. 10.5 µl were transferred to a new tube and supplemented with 1 µl 2S rRNA antisense DNA-oligo (100 µM). 2S rRNA antisense DNA oligonucleotides were annealed (5 min 75 °C; 15 min 37 °C; 15 min 25 °C) to block short and abundant 2S rRNA during 5'adapter ligation (Wickersheim and Blumenstiel 2013). Using pilot-PCR the minimal cycle number for the preparative smallRNA library was determined to 16-18 cycles. The amplicons were gel-purified and size-selected prior to sequencing on NextSeq500.  
attributes.tag: GEO Accession
attributes.value: GSM2975536