Small RNA-Seq of Drosophila melanogaster : control strain
Total RNA from 1st instar larvae was mixed with a very diluted solution of spike-in oligos (P-UGAGAUCAUUCGCAAAGCUGAUU and P-UCAAAAUAUUCGCAAAGCUGAUU; 5*10^(-22) mol per microgram of larval RNA; this concentration turned out to be too small for quantification in our setting). Small RNA-Seq library was prepared by the MGX sequencing facility (https://www.mgx.cnrs.fr/) using the NEXTflex Small RNA-Seq Kit v3 (Bioo Scientific). 5 adapter: GUUCAGAGUUCUACAGUCCGACGAUCNNNN; 3 adapter: NNNNTGGAATTCTCGGGTGCAAGG; RT primer: GCCTTGGCACCCGAGAATTCCA; barcode primer (P7): CAAGCAGAAGACGGCATACGAGATXXXXXXXXGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA and barcode primer (P5): AATGATACGGCGACCACCGAGATCTACACXXXXXXXXACACGTTCAGAGTTCTACAGTCCGA (with XXXXXXXX being the reverse-complement of the NEXTflex index). For this library, the index was: TTCAGGAG+TTAAGCTA. PCR products were size-selected on a PippinHT with a 3% cassette (selected size range: 137-183 bp). All 6 libraries were pooled in an equimolar fashion (after quantification of each library using Fragment Analyzer kit "High Sensitivity NGS" and qPCR (Roche), then seeded in the flow cell at 300 pM. Fastq output files were de-multiplexed using the bcl2fastq software (v 2.20.0.422).