RNA-seq of Placental Decidua of RPL
The RNA was fragmented using divalent cations under elevated temperature. Next, the cDNA is synthesized using Reverse transcriptase and random hexamers in a first strand synthesis reaction. Subsequently, the cDNA is converted to double stranded cDNA where Uracil is added instead of Thymine. The strand specificity is preserved by a USER enzyme-based digestion of second strand thereby leaving one functional strand which maps to the DNA strand which it was transcribed from. The USER digested single strand molecules were enriched and indexed in a limited cycle PCR followed by AMPure bead purification to create final cDNA library for sequencing