Project description:RBP9-TAP was expressed in bloodstream T. brucei and bound mRNA was pulled down using IgG beads and eluted via TEV-protease cleavage. rRNA was depleted using RNAseH and rRNA hybridizing oligos.
Project description:The first 4 samples belong to the RNA-IP using in situ TAP tagged ZC3H30 in procyclic (insect) form of the parasite T. brucei Lister 427, 2 samples are Elu or eluate, and 2 are FL or flowthrough (unbound) sample. The other 8 samples are also from procyclic cells. 4 samples belong to DKO(ZC3H30 gene double knockout), 2 are non-stressed and 2 are heat shocked samples; the rest 4 samples are DKO-ectopic (ZC3H30 double knockouts, expressing, ectopic copy of ZC3H30) 2 are non-stressed and 2 are heat shocked samples. Heat Shock experiment was done at 39 degree Celsius.
Project description:Pull-down of endogenous N-terminally TAP-tagged ERBP1 (Tb927.10.14150). RNA of unbound and eluted samples was purified, rRNA of unbound sample was depleted, and RNA was analysed by RNA-Seq.