GSM1208969: second_trimester_euploid_female2 [RNA-Seq]; Homo sapiens; RNA-Seq
RNA was extracted from 10 mL amniotic fluid supernatant (centrifugation at 350g, 4C, 10, min).All samples were processed using the Qiagen Circulating Nucleic Acid kit with an on-column DNase digestion step to remove genomic DNA. The RNA was then purified and concentrated with the RNeasy MinElute Clean up kit and eluted in RNasefree water. RNA was converted to cDNA and amplified using the Ovation RNA-Seq System V2 and then purified with the MinElute Reaction Cleanup Kit. Library preparation was undertaken on 500 ng cDNA according to standard practice with the alterations described: Fragmentation was not undertaken due to the already degraded and fragmented nature of cffRNA. In light of the small amount of starting material, a bead-based purification (AmPure XP Beads, Agencourt, Beckman Coulter, USA) was used in lieu of a gel-based size selection for the purpose of library clean up. Bioanalysis was performed to ensure the absence of primer-dimer peaks as well as determine the approximate insert size. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Illumina Hi-Seq 2000 following the manufacturer's protocols. One paired-end indexed library was sequenced per sample to a length of 50 nucleotides per mate.
GEO
GSM1208969