Analysis of yeast trimethylguanosine-capped RNAs by midwestern blotting.

Immuno-detection by 'Midwestern' blotting provides a simple way to identify trimethylguanosine (TMG) capped RNAs. With this technique, over 20 bands are observed when total cellular RNA from Saccharomyces cerevisiae is transferred to a nylon membrane and probed with anti-TMG antibodies. Most, if not all, species known to contain a TMG cap are detected by this method. Only TMG-capped RNAs are detected on Midwestern blots unlike anti-TMG immunoprecipitates. Midwestern blotting is a useful alternative to immunoprecipitation and Northern analysis and may prove to be a better method for determining the relative abundance of capped RNAs. The blots can be reprobed multiple times with labeled antisense oligonucleotides to determine the identity of any TMG-capped species for which the primary sequence or a clone is available. This dual detection capability provides a powerful tool for the analysis of TMG-capped snRNAs and snoRNAs.