Determination of the Identity and Organization of Messenger RNAs Within DNA

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  • Department of Biological Chemistry, Harvard Medical School, 25 Shattuck Street, Boston, Massachusetts 02115

We outline methods for identifying DNAs containing sequences complementary to specific mRNAs, and also provide a number of complementary approaches for mapping the arrangement of mRNAs along the DNA. These methods, together with S1 nuclease mapping and the direct visualization of R-loops with the electron microscope, provide a comprehensive approach to defining the architecture of mRNAs coding for specific polypeptides and the arrangement of RNA transcripts along the genome. This detailed cartographic information can then be used to study the steps in the processing of mature mRNAs and determine the modes by which the expression of specific genes is regulated.