RNA-Seq of Homo sapiens: female adult placenta
Briefly, 30 mg of tissue was disrupted in tubes with 5 mm metal beads under low temperature using two 2-minute cycles with 30 oscillations/minute in a TissueLyser LT (Qiagen) instrument. Tissue powder was resuspended in RTL buffer containing 10 l/ml -mercaptoethanol. A TRIzol reagent (Life Technologies, Rockville, MD) method was used and chloroform extraction performed prior to RNA purification using an RNeasy mini kit per the instruction manual. The quality and yield of the RNA samples were determined on a bioanalyzer platform using a total RNA nanoassay for eukaryotes (Agilent Technologies, Santa Clara, USA). Transcriptome libraries were prepared using the TruSeq Stranded Total RNA LT kit with Ribo-zero (Illumina, San Diego, USA). The 12 libraries were pooled together and sequenced in one lane of high-output 75 paired-end reads on an Illumina Next-Seq 500 sequencer.