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29

Shg1p associated RNAs


ABSTRACT: RNA co-immunopurification with TAP-tagged Shg1p (a component of the Set1 histone methyltransferase complex, SET1C) from Saccharomyces cerevisiae. An untagged (BY4741) served as a control (Gerber et al. 2004). Cells were grown to mid-log phase in rich media and harvested by centrifugation. TAP-tagged Shg1 was affinity-purified from cell-free extracts with IgG sepharose and eluted with TEV protease. RNA was isolated from extract (=input) and from the purified protein sample with the RNAeasy Kit (Qiagen). RNAs were reverse transcribed using a mixture of oligo-dT and random nonamer oligos in the presence of amino-allyl dUTP/dNTP mixture. cDNAs were fluorescently labeled and hybridized on yeast oligo microarrays over night at 42 degrees in formamide-based hybridization buffer. For the method see experimental procedure for RNA IP (Gerber/Dichtl). Overall design: An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract.

INSTRUMENT(S): SMD Print_1381 Saccharomyces cerevisiae

ORGANISM(S): Saccharomyces cerevisiae  

SUBMITTER: Andre Gerber  

PROVIDER: GSE15247 | GEO | 2009-08-31

SECONDARY ACCESSION(S): PRJNA116537

REPOSITORIES: GEO

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