GSM2976218: FW5547_Spt21 deletion total RNA-seq biological replicate 1; Saccharomyces cerevisiae; RNA-Seq
Cell pellets were washed once with sterile water and snap-frozen in liquid nitrogen. Total RNA was extracted using the hot acid phenol protocol (acid phenol:chloroform:isoamyl alcohol 125:24:1) and TE-SDS buffer, then precipitated in ethanol with 0.3 M sodium acetate before re-suspension in DEPC-treated sterile water. Total RNA from yeast was incubated with rDNase (Machery-Nagel rDNase Set) and column purified (Machery-Nagel NucleoSpin RNA) prior to sequencing library preparation. For total RNA-Seq libraries, intact yeast total RNA was depleted of rRNA with the Illumina RiboZero Gold rRNA removal kit (yeast) (Illumina MRZY1324). 1 μg of yeast total RNA was used to generate strand-specific libraries for total RNA sequencing using the TruSeq Stranded mRNA Kit (Illumina 20020594) according to the manufacturer's instructions. Library quality control was performed using the Agilent 2200 TapeStation with D1000 screentape (Agilent Technologies). Each library was sequenced on the HiSeq 2500 platform using V4 chemistry (Illumina) and typically generated ~37 million 101 bp strand-specific paired-end reads per sample.
GEO
GSM2976218