Small RNA-Seq of Drosophila melanogaster : m2/+ strain
Total RNA from 1st instar larvae was mixed with a correctly-diluted solution of spike-in oligos (P-UGAGAUCAUUCGCAAAGCUGAUU and P-UCAAAAUAUUCGCAAAGCUGAUU; 1.5*10^(-16) mol per microgram of larval RNA; this concentration turned out to be suitable 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: CACATTAC+CACGCCGG. PCR products were size-selected on a PippinHT with a 3% cassette (selected size range: 137-183 bp). Library was seeded in the flow cell at 300 pM.