title: GSM8264762: Early embryo, tm2321, rep 2; Caenorhabditis elegans; miRNA-Seq
library_construction_protocol: Samples were washed with M9 buffer and then snap-frozen. TRI reagent was added to frozen worm pellets, which were then thawed and snap-frozen twice more to disrupt the embryonic cuticle. Phase separation was achieved with 1-bromo-3-chloropropane, and the aqueous phase was precipitated in isopropanol. Libraries were prepared from 2–10 µg total RNA essentially as described (Kleaveland, 2018), with the addition of spike-in RNA oligonucleotides in proportion to the total RNA content of each sample. A step-by-step protocol for sRNA-seq library preparation can be found at http://bartellab.wi.mit.edu/protocols.html.