GSM4910501: Ycg1Degron1 G1 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 cleaned up through column purification through the RNA Cleanup protocol within the RNeasy Mini Kit including DNase digestion (Qiagen) prior to sequencing library preparation. For total RNA-Seq libraries, the library was contructed with KAPA RNA HyperPrep kit without the riboerase treatment. The polyA RNA-seq libraries were contructed using the KAPA mRNA HyperPrep kit according to the manufacturer's instructions. Library quality control was performed using the Agilent 2200 TapeStation with D1000 screentape (Agilent Technologies). RNA sequencing was carried out on the Illumina HiSeq 4000 platform and typically generated approximately 18 million 76 bp strand-specific single-end reads per sample.