title: GSM1139042: Hybrid_RNASeq_01; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: Each of the replicated subcultures of BYxRM hybrid and BY+RM co-culture harvested at OD600=1.0 was used to extract total RNA by the hot acid phenol method (Kohrer and Domdey 1991) mRNA-seq library preparation used the Illumina TruSeq mRNA-seq Sample Prep kit with some modifications. Briefly, 5 μg of total RNA from each sample was used to purify for polyA-RNA, and the mRNA was fragmented by heat at 94℃ for 8 min. Double-stranded cDNA was synthesized by random priming, end repaired, and ligated to the Y-shaped TruSeq adaptors. Samples were cleaned by AMPure beads (Agencourt) and split into two halves, which were independently assembled into two PCR reactions, one using the PCR reagents provided in the Illumina kit, and the other using the KAPA PCR reagent (KAPA HiFi HotStart ReadyMix). All reactions went through 12 cycles of PCR amplifications, and were cleaned by AMPure beads to remove primer dimers. The purified products were quantified by Qubit (Invitrogen) and BioAnalyzer 2100 High Sensitivity DNA Assay (Agilent).
attributes.tag: GEO Accession
attributes.value: GSM1139042