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The inability of SI nuclease to detect mismatches of less than 4 contiguous nucleotides limits the usefulness of Berk & Sharp SI mapping for identifying small mutations in mRNA. This prompted Myers et al and Winter et al to develop new techniques utilizing RNase A to cleave at mismatches between a radlolabeled RNA probe and complementary RNA or DNA. These powerful techniques, however, can only be used with < uniformly labeled RNA probes. We have devel•* •* « oped a modified method which allows the use 8 $ J of 3' or 5' end-labeled DNA probes for the Z z a: detection of mutations in mRNA. Such end<* labeled probes may simplify the localization =<=L o of mutations in mRNA. This method is "~ m •extremely simple and involves standard Berk & ,_ + + i Sharp SI hybridization conditions followed by </> w </j w t h e addition of 75 units of SI nuclease + 1 ug of RNase A to a standard SI digestion buffer. The figure shows such a study where RNA from a mutant g-globin gene with a single 9 • base substitution at codon 17 (A to T) is

Detecting small mutations in expressed genes by a combination of S1 nuclease and RNase A