Dataset features


Application: RNA-seq analysis
Number of samples: 18
Release date: Mar 30 2018
Last update date: May 2 2018
Access: Public
Dataset link C. elegans FBF iCLIP from spermatogenic and oogenic germlines

Experimental Protocol

We generated FBF-1 and FBF-2 iCLIP datasets from animals with somatic tissues of the same gender but germline tissues of opposite gender. For each FBF, we used an N-terminally FLAG-tagged single copy fbf transgene in a strain lacking the endogenous gene (e.g. tagged FLAG::FBF-1 in an fbf-1 null mutant ). Adult hermaphrodites with a sexually transformed germline that are strictly spermatogenic were obtained using a temperature sensitive gain of function mutation in the fem-3 gene (Barton and Kimble 1987). These fem-3 (ts, gf) mutants are spermatogenic at restrictive temperature (25°C). We therefore introduced FLAG-tagged FBF-1 or FBF-2 into the fem-3(ts,gf) mutant, again removing the corresponding endogenous fbf gene in each strain. We then performed FBF iCLIP from adults that were either oogenic or spermatogenic, both raised at 25 °C. For each strain (each FBF, each germline gender), we processed three biological replicates. Three negative control datasets were produced for each germline gender in parallel by omitting the FLAG antibody.