GSM2800847: Act88F female -ox; Drosophila melanogaster; ncRNA-Seq
Total RNA was extracted from 2-5 days old adult female flies collected in Trizol using a phenol-chloroform-based standard protocol. Small RNA sequencing libraries were constructed from an input of at least 80ug total RNA. Small RNA libraries were constructed and sequenced as described in Ameres et al. (2010) with some modifications. For miRNA deep sequencing datasets, 18-30 nt long RNAs were size selected. Subsequent 2S rRNA depletion was performed with Dynabeads MyOne streptavidin C1 beads (Invitrogen) as described in Seitz et al. (2008). Small RNAs were ligated to 3' and 5' barcode adapters containing 4 random nucleotides at their ends to minimize ligation bias (Jayaprakash et al., 2011). Subsequent reverse transcription was performed with SuperScript III Reverse Transcriptase (Invitrogen) and cDNA samples were PCR amplified with the KAPA Real-Time Library Amplification Kit (Peqlab). Amplified cDNA was agarose gel purified and sequenced in an Illumina HiSeq2500 instrument.
GEO
GSM2800847