Ectocarpus mature gametophyte small RNAs- Female 2
Small RNAs were isolated and prepared for sequencing by Fasteris (Plan-les-Ouates, Switzerland). Between four and 12 micrograms of total RNA was extracted for each replicate using the Qiagen Mini kit. RNA was separated on a polyacrylamide gel and the 15 to 30 nucleotide fraction isolated by excision. Addition of single-stranded adapters and PCR amplification was carried out using the DGE-Small RNA kit (Illumina, San Diego, USA) and small RNAs were sequenced on a HiSeq 2000 (Illumina).