Abstract
The application of molecular biological techniques to immunological studies is based in large part on the manipulation of nucleic acids--RNA, DNA, and oligonucleotides. This unit offers very basic procedures that are used in a number of protocols later in the chapter. In particular, preparation of nucleoside triphosphates--the substrates for almost all synthetic reactions using nucleic acids--is described; the relevant radioisotopes, (32)P, (35)S, and (3)H are discussed; and methods for quantitating radioisotope and biotin incorporation into nucleic acids are presented. Finally, column chromatographic (gel filtration) procedures for separating labeled DNA from synthetic precursors are described.
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