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. 2007 Nov 15;26(24):4996–5006. doi: 10.1038/sj.emboj.7601921

Figure 2.

Figure 2

Cryptic transcripts can be detected from regions of repressed chromatin in strains lacking TRAMP activity. (A) Northern analysis of TRAMP mutants grown in YPD at 25°C. RNA is resolved on a 1.2% agarose glyoxal gel (three upper panels) or a 6% acrylamide 7 M urea gel (three lower panels). Probes are (top to bottom) YEL074W, NTS1 HpaI, TSA1, CEN3, MAT and TSA1. (B) Schematic representations of probed regions. (C) 5′ RACE PCR was performed on wild-type and trf4Δ RNA, with a primer located 138 bp from the 3′ end of the annotated YEL077C ORF. DNA products were stained with SYBR Safe. The indicated band was excised, cloned and sequenced. (D) Northern analysis of TRAMP and exosome mutants probed with YEL074W and PGK1. Strains were grown at 25°C on YPD except trf4Δ GAL-trf5, which was grown on YPGal and shifted to YPD for 18 h prior to RNA extraction.