In this article, we describe a new procedure to map 5′ ends of RNAs. The procedure consists in the use of specific RNase H digestion of a hybrid formed by the RNA and a complementary DNA oligonucleotide. Northern blot hybridization of the resulting RNA fragment allows an accurate measurement of its length. Although we generally use this procedure as a control of previously performed primer extension analyses, the absence of nonspecific bands, which often occur in primer extensions on RNA templates with extended secondary structures, suggests that our method may be preferable when these difficult templates are analyzed.
BELLAVIA D, FORTE GI, SISINO G, SISINO G, BARBIERI, R. (2008). A new method for the mapping of 5' ends of RNAs. ANALYTICAL BIOCHEMISTRY, 378(1), 113-114.
A new method for the mapping of 5' ends of RNAs
BELLAVIA, Daniele;FORTE, Giusi Irma;BARBIERI, Rainero
2008-01-01
Abstract
In this article, we describe a new procedure to map 5′ ends of RNAs. The procedure consists in the use of specific RNase H digestion of a hybrid formed by the RNA and a complementary DNA oligonucleotide. Northern blot hybridization of the resulting RNA fragment allows an accurate measurement of its length. Although we generally use this procedure as a control of previously performed primer extension analyses, the absence of nonspecific bands, which often occur in primer extensions on RNA templates with extended secondary structures, suggests that our method may be preferable when these difficult templates are analyzed.
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