Figure 2.

Validated putative miRNAs. (A) Sequences for both mature miRNA and predicted pre-miRNA. Copy number refers to the occurrences of the mature sequences in the E15.5 whole brain small RNA sequencing analysis. (B) RNAfold prediction of the stemloop hairpin structure. The colours in the vertical bar denote the base-pairing probability between two nucleotides within the structure. The black line located next to the hairpin structure denotes the position of the small RNA within the pre-miRNA. (C) Small RNA northern analysis using radiolabeled oligonucleotide probes. 'p' and 'm' refer to pre-miRNA and mature miRNA, respectively. Four independent small RNA northern blots were used to validate the putative miRNA. After hybridization and washing steps, mm_br_e15_1181 blot was exposed to phosphor screen for 1 day whereas the other 3 blots for mm_br_e15_279920, mm_br_e15_96719 and mm_br_e15_294354 were exposed for 8 days. (D) Mapping of the mature miRNA to the mouse genome and other corresponding features such as RefSeq genes, miRNAs from miRBase, mouse ESTs, mammalian conservation information and repeating elements.