Identifying expression of new small RNAs by microarrays

Although a large number of small RNAs (sRNAs) have been discovered, it is very likely that the screens conducted so far have not reached saturation. Recently, many methods for predicting and identifying new sRNAs have been developed. However, it remains unclear what the total number of sRNAs within a genome is and how many types of sRNAs exist in plants and animals. In this article, combined methods of dynamic programming prediction, enrichment of sRNAs, and microarray analysis are developed to screen and evaluate a new class of sRNAs from introns of human, protein-encoding genes. The methods used by our laboratories to design capture probes and label enriched small RNAs are thoroughly described here. The microarray results show that our modified technologies are useful to enhance sensitivity and specificity of arrays, identify expression patterns within different cells, and discover differential expression of sRNAs during the differentiation process of bone marrow stem cells. Accordingly, the combination of computational prediction and microarray analysis may be a feasible and practical approach for profiling studies of both known and predicted small RNAs.