Detection of nonpolyadenylated RNAs. (A) I-tailing of total yeast RNA. Yeast RNA treated with (+) or without (−) Cid1 and ITP for I-tailing were run on a 6% denaturing polyacrylamide gel and stained with SYBR Gold. Slight shift up of 18 and 25S rRNAs indicates addition of poly(I). Disappearance of 5.8S rRNA is consistent with its acquisition of a long heterogeneous tail (See Fig. 2). (B) Genome browser view of the rRNA genes of S. cerevisiae showing detection of rRNA by different library preparation methods. Top line is poly(A) selected RNA using the direct oligoT capture method, second line is poly(A) selected and I-tailed RNA captured with the oligoC adapter, third line is total RNA I-tailed and captured with the oligoC adapter, fourth line is Ribominus rRNA depleted total RNA I-tailed and captured with the oligoC adapter. (C) Scatter plot showing robust detection of nonpolyadenylated noncoding RNAs by I-tailing and capture with the oligoC adapter.
