IMPROVED COLUMN FOR DNA SEPARATION BY MATCHED ION POLYNUCLEOTIDE CHROMATOGRAPHY

摘要

An improved separation column and method for separating a mixture of double stranded DNA fragments by Matched Ion Polynucleotide Chromatography. The cylindrical column has an ID greater than about 5 mm and contains polymer beads. The beads have an average diameter of 1 to 100 microns and are unsubstituted polymer beads or are polymer beads substituted with a hydrocarbon moiety having from 1 to 1,000,000 carbons. The preferred beads a re characterized by being substantially free from multivalent cations which are free to bind with DNA. The improved column provides enhanced separation of D NA fragments with sizes ranging from about 100 to 20,000 base pairs. The column also provides enhanced separation of heteroduplex and homoduplex DNA molecul es in a mutation detection procedure in which the chromatography is performed under conditions effecting partial denaturation of DNA at a site of mismatch ed base pairs.