GSE160993
Genetic and pharmacological evidence for kinetic competition between alternative poly(A) sites in yeast - 4tU cordycepin treatment
mPAT approach using an anchored oligo-dT (TV12VN) primer to determine changes to 3'UTR length in yeast upon cordycepin treatment and new transcription using 4-thiouracil (4tU) labelling of RNA. This appears as a T to C conversion in sequencing reads. Overall design: BY4741 yeast cells were grown at 30ºC until an OD600 of 0.6 was reached then 20µg/ml cordycepin and 500µM 4tU or equivalent volumes of DMSO added for 0, 20 or 40 minutes before harvesting. Samples were analysed via mPAT with an anchored oligo-dT primer using the Illumina MiSeq platform.