Validation of Yersinia pestis sRNAs with Nonradioactive-Labeled Northern Blotting
Despite the modification of several existing techniques to detect low-level sRNAs, such as the nuclease protection assay and reverse transcription-PCR, the classic Northern blotting technique has remained irreplaceable. The classic Northern blotting technique is a relatively simple and straightforward procedure that allows the visualization and rough quantification of cellular levels of specific RNA species and their processing intermediates relative to endogenous RNA standards. A Northern blotting analysis of small RNAs (sRNAs) involves the size fractionation of the RNAs in a gel, followed by their transfer onto a solid medium (membrane) to maintain the relative positions of the RNA molecules. The transferred molecules are tightly immobilized, usually with UV cross-linking. A labeled probe complementary to the mRNA of interest is hybridized to the membrane. The signal generated by the probe can be used to determine the size and abundance of the target RNA.
Key wordsYersinia pestis sRNA Validation Northern blot
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