Illumina sequencing of Saccharomyces cerevisiae transcript fragment library
Total RNA was extracted from Ribopure Yeast kit (Ambion) and treated for 30 min at 37C with RNAse free DNase I (Ambion). Poly(A) RNA was purified with Micro Poly(A) Purist Kit according to the manufacturer's instructions (Ambion), and used to generate double stranded cDNA by reverse transcription with either random hexamers or oligo(dT) primers. The double stranded cDNA was fragmented and subjected to high throughput Illumina sequencing in which 35 bp of sequence was determined from the fragment ends.