Investigation of the stereoconfiguration 5' of incorporated uridines.

PhosphorImager analysis of an analytical 25% polyacrylamide gel of nuclease P1 (lanes 1,3,6,9) and snake venom phosphodiesterase (PDE) (lanes 2,4,7,10) digestions. (Lanes 1,2): Digested RNA uniformly labeled with [a-32P]UTP. (Lanes 3,4): Digested RNA uniformly labeled with [a-35S]UTP. (Lanes 5,8): Undigested 5' or 3' fragment, respectively, labeled by incubation with mitochondrial extract and [a-35S]UTP. (Lanes 6,7): Digested 5' fragment, labeled by incubation with mitochondrial extract and [a-35S]UTP. (Lanes 9,10): Digested 3' fragment, labeled by incubation with mitochondrial extract and [a-35S]UTP. The positions of the undigested 5' and 3' fragments as well as the mono- and di-nucleotide cleavage products are indicated. 32P and 35S radioisotopes are shown as *p and *S, respectively. The relative numbers of added U's per site can be estimated from the nuclease P1 digestion lanes after the appropriate normalization for the number of labeled U residues per band.

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