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. 2011 Dec;17(12):2152–2164. doi: 10.1261/rna.028118.111

FIGURE 1.

FIGURE 1.

Identification of a small RNA and its precursor in Giardia. (A) Sequence of the 972-nt ORF from which the small RNA and its precursor are derived. The small RNA is boxed in red, while the precursor is underlined. (B) Northern blot analysis of total RNA (left panel) showed the presence of an ∼1000-nt RNA that corresponds to the GL50803_92663 ORF transcript, suggesting that it is expressed. An ∼100-nt band and an ∼70-nt band that correspond to bands seen on the Northern blot of size-fractionated small RNA (<200 nt) were also observed. Additional bands may represent GlDcr intermediate products or degradation. Northern blot analysis of size-fractionated small RNA (<200 nt) identified an ∼30-nt band corresponding to the small RNA and an ∼100-nt band that may correspond to its precursor (right panel). Additional faint bands were also observed at ∼60 nt and ∼70 nt. (C) Primer extension of size-fractionated (<200 nt) RNA from Giardia trophozoites identified a 94-nt fragment and a 26-nt band corresponding to the small RNA. Additional bands shown by color-coded arrows have their corresponding locations indicated on the pre-miR4 secondary structure below. (D) MFOLD secondary structure of the 94-nt RNA identified by primer extension. The small RNA is indicated in red.