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- W1005026897 abstract "Publisher Summary This chapter describes procedures for the isolation, purification, and analysis of the branched oligonucleotides obtained either from RNA labeled in vivo or from intermediates formed in splicing extracts from a specific 32 P-labeled pre-mRNA. Success depends partly on the concentration of branched intermediates in the sample and on the levels of other unidentified components that may cochromatograph with branch points. Branch points, recovered from the digests of RNA lariats purified from nuclear-splicing extracts do not require extensive purification. Secondary analysis is simplified by the removal of the excess mononucleotides generated by nuclease digestion. A DNA template is prepared by inserting—at appropriate restriction sites of a transcriptional cloning vector—a complete intron flanked by exons. The isolation and purification of branched oligonucleotides from mixed RNA populations usually require more elaborate purification schemes, because of the presence in nuclease digests of pre-mRNAs of modified structures, such as caps, methylated nucleotides, and other unidentified components, as well as pNps, which may copurify with branched oligonucleotides because of similarity in net charge." @default.
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- W1005026897 date "1989-01-01" @default.
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- W1005026897 title "[15] Isolation and characterization of branched oligonucleotides from RNA" @default.
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- W1005026897 doi "https://doi.org/10.1016/0076-6879(89)80101-6" @default.
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