IDENTIFICATION OF NUCLEIC-ACIDS AND OLIGONUCLEOTIDES BY CAPILLARY ZONE ELECTROPHORESIS WITH THE 4-MARKER TECHNIQUE

The paper describes the improvement brought by the four marker techniq ue to the repeatability of the separation of macromolecules [nicotinam ide nucleotides (NADPH and NADH), one adenine compound (ATP), two olig onucleotides (135-oligomer and 84-primer containing 135 and 20 bases, respectively) and N2-5'-de oxyquanosine-benzo[a]pyrene-diolepoxide (dG 5'BaDE)] by capillary zone electrophoresis with UV detection at the wa velength of 220 nm. Optimized conditions [3-(cyclohexylamino)-1-propan esulphonic acid as the organic electrolyte solution with pH of 11.2, a nd concentration of 50 mM] were used for the separation. The counter-i on in the system was sodium. The macromolecules could be separated wit hin 20 minutes. The four marker compounds were (-)mandelic acid, meso- 2,3-diphenylsuccinic acid, 1,2-phenylenediacetic acid and o-phthalic a cid, which were used for the calculations of the mobilities for the an alytes. The identification of the macromolecules was confirmed with th e coefficients of identification. The reliability of the identificatio n was increased up to 75-fold relative to the use of the absolute migr ation times.