RNA-seq of UTEX2973, mixture sample, random library
The RNA was treated by the Ribo-Zero rRNA Removal Kit for ribosomal RNA (rRNA) depletion. The rRNA depleted RNA samples were fragmented using ultrasound (4 pulses of 30 s each at 4 C). Then, an oligonucleotide adapter was ligated to the 3' end of the RNA molecules. First-strand cDNA synthesis was performed using M-MLV reverse transcriptase with the 3'-adapter as a primer. The first-strand cDNA was purified, ligated with the 5' Illumina TruSeq sequencing adapter, and amplified by PCR (11 cycles).