GSM6398555: MS 2; Sus scrofa; miRNA-Seq
In brief, dilute plasma ﬂuid with an equal volume of PBS and centrifuge 30 min at 2,000 × g, 4◦C, to remove cell. Then, carefully transfer supernatant to ultracentrifuge tubes or bottles without pellet contamination, centrifuge 45 min at 12,000 × g, 4◦C. Next, filter the suspension through a 0.22-µm ﬁlter, and collect in a fresh ultracentrifuge tube or bottle, centrifuge 2h at 160,000 × g, 4◦C. Pour off the supernatant. Resuspend the pellet in 250µl PBS and Store at −80◦C until use. Three MS and threeOS piglets were used for library construction. Approximately 1 μg of total RNA was used to prepare the small RNA library according to the protocol of TruSeq Small RNA Sample Prep Kits (Illumina). Single-end sequencing (36 bp) on an Illumina Hiseq 2500 at the LC-BIO (Hangzhou, China) was performed following the recommended protocol.