title: GSM1261669: RNA-seq, WT C. albicans; Candida albicans; RNA-Seq
library_construction_protocol: Ribosome profiling: Cells from log-phase cultures were flash frozen in liquid nitrogen. To guard against post-lysis translation in the ribosome-profiling sample, we supplemented the lysis buffer with 50µg/ml GMPPNP and 10µg/ml Blasticidin S (Invivogen). Cells were lysed via mixermilling, and lystates were cleared via centrifugation and then treated with RNase I before loading onto a sucrose gradient and ultracentrifugation. Monosomes were collected, and ribosome-protected mRNAs were extracted with acid phenol and chloroform. RNA-Seq: Cells from log-phase cultures were flash frozen in liquid nitrogen, and mRNA was isolated using acid phenol and chloroform. mRNAs were selected using oligo-dT beads (Invitrogen) and then alkaline fragmented. Ribosome footprints (FPs) and mRNAs were size-selected on TBE-Urea gels and dephosphorylated prior to ligation with a common linker, rRNA subtraction (FP sample only), and RT on both samples. DNAs were circularized and then amplified by PCR to yield fragments suitable for deep sequencing.
attributes.tag: GEO Accession
attributes.value: GSM1261669