A new method for the mapping of 5' ends of RNAs.
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
Daniele
D
Dipartimento di Biologia Cellulare e dello Sviluppo, Università di Palermo, Palermo 90128, Italy.
Forte
Giusi Irma
GI
Sisino
Giorgia
G
Sisino
Giada
G
Barbieri
Rainer
R
D000818:Animals; D015152:Blotting, Northern / Q000379:methods*; D012313:RNA / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism; D012335:RNA, Ribosomal / Q000032:analysis / Q000235:genetics; D016914:Ribonuclease H / Q000378:metabolism; D012617:Sea Urchins / Q000235:genetics; D013698:Templates, Genetic
D012335:RNA, Ribosomal; D012313:RNA; D016914:Ribonuclease H