GSM8767728: PL296_v; Homo sapiens; ncRNA-Seq
Sorted cells were isolated from fresh chorionic villi using FACS as described in GSE159526. Sorted cells and chorionic villi samples were extracted using the ZYMO Quick-RNA MiniPrep kit according to manufacturer's instructions or using TRIzol (ThermoFisher Scientific, USA). 5.8S rRNA was masked during adapter ligation by a complementary oligonucleotide, while miRNAs underwent 3′ adapter ligation, digestion of unligated adapters, and 5′ adapter ligation sequentially. The 5′ adapter contains a unique molecular identifier (UMI), followed by two nucleotides (CA) prior to the miRNA sequence. Upon conversion to cDNA by reverse transcription, the cDNA was amplified by PCR to include the sequences required for Illumina cluster generation and sample indexing developed by the GSC. The number of PCR cycles was dependent on the amount of input RNA; samples with low and high input were subjected to 20 and 15 cycles of PCR, respectively. Quality of libraries was assessed using the Agilent Bioanalyzer 2100 HS DNA chip Assay, (Agilent, USA)
GEO
GSM8767728_r1