Figure 2

RNA of cells lysed by French press is degraded. Strains RJY358 (wt, untagged), RJY933 (Pbp2p-TAP) and RJY929 (Nrp1p-TAP) were lysed in a French Press and samples from crude lysate, supernatant of 1200 × g (S1), 20000 × g (S20), 200000 × g (S200) spins and pellet of 200000 × g (P200) spin were phenol extracted. 8 μg of the extracted RNA were loaded onto 1.2% agarose-formaldehyde gels and blotted onto nylon membranes. 8 μg of total RNA (P/C lysis RNA) from the same strains prepared with a phenol extraction method [18] were loaded in parallel as control for intact RNAs. After methylene blue staining, the membranes were hybridized with a probe against PDA1 mRNA. The positions of the 18S and 25S ribosomal RNAs in the methylene blue staining and the PDA1 mRNA hybridization signal are indicated. The nature of the third band that appears in the methylene blue staining on top of the 25S rRNA is unknown.