GSM5001824: MX1303-C5JC4ACXX-3-TAGGAT; Homo sapiens; ncRNA-Seq
Thirty milligram samples were taken from the chorionic villi on the fetal-facing side of the placenta. TRIzol was used to extract total RNA. Sequencing was performed using the standard ribodepleted strand-specific RNA sequencing protocol of Canada's Michael Smith Genome Sciences Centre in Vancouver, in line with The Cancer Genome Atlas miRNA sequencing protocol (Chu et al., 2015, PMID: 26271990). Briefly, after adaptor ligation, amplification was achieved through 15 cycles of PCR (98˚C for 15s, 62˚C for 30s, 72˚C for 15s), concluded by 5 minutes at 72˚C. Gel extraction from a 3% MetaPhor Agarose gel was used to select for RNAs shorter than 200 nucleotides, and hybridization was used to deplete the rRNA fraction. RNA quality was confirmed on an Agilent Bioanalyzer 2100 (Agilent, USA). Sequencing was performed on an Illumina HiSeq 2500.