3 protocols
normalization data transformation protocol
Illumina Cluster Station and Illumina HiSeq™ 2000 instrument used for basecalling. Raw sequences have 3' adaptor fragments as well as a few low-quality sequences and several types of impurities. Raw sequences are transformed into Clean Tags after certain steps of data-processing. A virtual libraries containing all the possible CATG+17 bases length sequences of the reference gene sequences.All clean tags were mapped to the reference sequences and only 1bp mismatch is considered. Clean tags mapped to reference sequences from multiple genes were filtered. Remainder clean tags were designed as unambiguous clean tags. The number of unambiguous clean tags for each gene was calculated and then normalized to TPM (number of transcripts per million clean tags). ('t Hoen, Ariyurek, et al. 2008; Morrissy, Morin, et al. 2009). Supplementary_files_format_and_content: tab-delimited text files include tag sequences,tag number and TPM values for each strain.
growth protocol
Yeast-like cells were cultivated in 2% potato dextrose broth (PDB) at 20 degree into their exponential growth phase.
nucleic acid library construction protocol
Total RNA was extracted from cells using pBIOZOL Plant Total RNA Extraction Reagent, according to the manufacturer’s protocol (BioFlux). The isolated RNA was treated with RNeasy plant mini kit to remove potential genomic DNA contamination, according to the manufacturer’s protocol (QIAGEN, Germany). RNA integrity and concentration were evaluated using an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA). RNA (6 μg) from each of the three strains was submitted to BGI-Shenzhen (Shenzhen, China) for library construction and sequencing. Sequence tag preparation was carried out using the Illumina Gene Expression Sample Prep Kit according to the manufacturer’s protocol.