GSM2147999: Dhh1-DQAD-OE mRNA-Seq; Saccharomyces cerevisiae; RNA-Seq
For ribosome footprint profiling, lysates were clarified and monosomes were isolated by sucrose gradient centrifugation after RNase treatment. RNA was extracted using SDS/hot phenol/chloroform. For mRNA-Seq, total RNA was extracted from frozen cells by using SDS/hot phenol/chloroform. For ribosome footprint profiling, fragments ranging from (15/25)-34 nt were gel purified and rRNA was depleted from this pool through Ribo-Zero Gold (Yeast) treatment. Upon dephosphorylation of sample RNA, IDT miRNA Cloning Linker 1 was ligated and this product was once again gel purified. This was followed by reverse transcription as well as circularization of cDNAs. PCR amplification of this library was then sent for sequencing. For mRNA-Seq, rRNA was depleted through Ribo-Zero Gold (Yeast) treatment and the remaining RNA was randomly fragmented with light base treatment. Fragments of 20-40 nt were prepared using the above protocol.
GEO Accession
GSM2147999