171_days_postpartum_Endometrium_Intercaruncular
Stranded RNA-seq libraries were constructed using the poly-A enrichment method with the Illumina Stranded mRNA Prep (Illumina, San Diego, CA). Briefly, the poly-A containing mRNA was selected, purified, and fragmented into small pieces. The first-strand cDNA was synthesized using reverse transcriptase and random primers, followed by second-strand cDNA synthesis marked to allow strandedness. The fragments were adenylated to prevent the 3 ends from ligating to each other. Pre-index anchors were ligated to the ends of the cDNA, followed by purification of adapter-ligated fragments. Anchor-ligated DNA fragments were amplified, and indexes added, yielding a dual-indexed library ready for sequencing. Each purified library was assessed using the Fragment Analyzer, quantified with a fluorometer (Qubit), and sequenced on NovaSeq 6000 sequencer (Illumina) using a single NovaSeq S2-PE100 flow cell with an average read count of 50 million paired reads per sample.