GSM9116770: lsm1-ideg strain T60, repl. 2; Saccharomyces cerevisiae; RNA-Seq
Total RNA was extracted with hot phenol from yeast cells and precipitated in the presence of ammonium acetate and ethanol. After additional washes in 70% ethanol, RNA was recovered in pure water. The Novogene company performed the sequencing, using poly(A) selection and strand specific RNA sequencing. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers. Then the second strand cDNA was synthesized using dUTP, instead of dTTP. The directional library was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification.