GSM7927373: blood_PRYA146; Homo sapiens; RNA-Seq
Total RNA was extracted from placenta samples using the Qiagen Rneasy Mini kit cat #74104 and from blood samples using the Ribo kit Invitrogen AM1928 For each sample 100 ng of total RNA were used for library preparation with the Illumina RNA Prep with Enrichment (L) Tagmentation kit (Illumina, cat # 20040536) following manufacturer's instructions. Briefly, after the first and second strand cDNA synthesis, double-stranded DNA was subject to tagmentation, where the Transposomes simultaneously tagmented and added adapters to the double-stranded cDNA. After tagmentation, the fragments were purified and amplified to add index adapter sequences (IDT for Illumina DNA/RNA UD Indexes Set B, cat # 20027214) for dual indexing and P7 and P5 sequences for clustering. 200 ng of library were hybridized to enrichment oligos (Illumina Exome Panel cat # 20020183) with an overnight incubation at 58°C. The enriched libraries were amplified with 14 PCR cycles and cleaned up with Agencourt AMPure XP beads (Beckman Coulter cat # A63880). The libraries quality control was performed using the Qubit dsDNA BR assay kit (Thermo Fisher Scientific, cat # Q32850) and the Agilent D1000 ScreenTape assay (Agilent, cat # 5067-5582). The libraries were pooled after quality control followed by sequencing on a NovaSeq 6000 SP lane as paired-end 100 bases.