title: GSM2482043: S. acidocaldarius L7Ae Co-IP small RNA; Sulfolobus acidocaldarius; RNA-Seq
library_construction_protocol: Sample 1: Cells were lysed by homogenization and small RNAs were isolated according to the mirVana miRNA Isolation Kit (Ambion). The small RNA isolate was used for cDNA library preparation. Samples 2 and 3: In order to obtain L7Ae-interacting RNAs, cells with a C-terminal Flag-HA tag on the single L7Ae gene (saci_1520) in S. acidocaldarius were lysed by a French Pressure Cell Press and the protein was purified by two consecutive immunoprecipitation steps using the Flag-HA tandem purification Kit (Sigma). The urea eluate contained co-immunoprecipitated RNA. One sample was fragmented by ZnCl2 (10 mMZnCl2, 20 mM Tris pH 6.8) and treated with T4 PNK to cure the RNA termini. The two RNA samples (L7Ae co-IP small RNA and fragmented RNA) were then used for cDNA library preparation. cDNA libraries were prepared according to the NEBNext Small RNA Library Prep Set for Illumina.
identifiers.namespace: GEO
identifiers.id: GSM2482043