GSM3594066: 30minusstrand; Saccharomyces cerevisiae; OTHER
Cells lyses in SamplePrep 6870 freezer mill (SPEX SamplePrep) were performed. Lysates were sonicated and ultracentrifugated. Solutions were incubated in Ni-NTA agarose nickel beads (Qiagen). Incubation with streptavidin beads and treatment with RNase was accomplished. Lysates were treated with Thermosensitive Alkaline Phosphatase (Promega), T4 PNK (New England Biolabs) and 32P γ-ATP. Adaptors were bound to RNA and then incubated with Proteinase K (New England Biolabs). RNA obtained was purified by acid phenol/chloroform extraction/ethanol precipitation. Reverse transcription, PCR amplification, cDNA multiplexing and gel extraction of cDNA libraries were accomplished.
GEO Accession
GSM3594066