RPC-5 column chromatography is a useful tool for the high resolution fractionation of single-stranded DNA or RNA oligonucleotides, double-stranded DNA restriction fragments, and the complementary strands of DNA restriction fragments. The technique can be used for fractionation of milligram quantities of nucleic acids. The order of fractionation is not always the same as produced by gel electrophoresis. Therefore, the sequential use of these two methods is more powerful than either method alone. In some cases RPC-5 column chromatography can completely resolve duplex restriction fragments of the same size. The chapter describes the technical details of the fractionation procedures. The practical laboratory methodology is emphasized. The future uses of RPC-5 are: the separation of DNAs that have small differences in their content of nonpaired nucleotides and the separation of certain protein-nucleic acid complexes (such as nucleosomes or transcription complexes) from naked DNA.
ASJC Scopus subject areas
- Molecular Biology