body9
Total RNA was extracted after thawing the tissues and electric homogenization using TRIzol followed by purification with the Zymo Quick-RNA MiniPrep kit. Libraries were prepared using the Genomics in house protocol at the Weizmann Institute of Sciences Genome Center for mRNA-seq. Briefly, the polyA fraction (mRNA) was enriched from 500 ng of total RNA followed by fragmentation and the generation of double-stranded cDNA. Then, end repair, A base addition, adapter ligation and PCR amplification steps were performed. Libraries were evaluated by Qubit (Thermo fisher scientific) and TapeStation (Agilent). Sequencing libraries were constructed with unique barcodes for each sample to allow multiplexing.  Around 18-24 million 127-bp Paired-End reads were sequenced per sample on an Illumina HiSeq High Output instrument.