Sequence-dependent DNA separation by anion-exchange high-performance liquid chromatography.
High-performance liquid chromatography (HPLC) system with a new nonporous anion-exchange resin, DNA-NPR, made it possible to rapidly separate DNA fragments up to 20 kbp with high resolution. In order to further characterize this chromatographic DNA separation system, we prepared a mixture of double-stranded DNAs of constant length carrying a fully degenerated 50-bp region and analyzed their chromatographic behavior on the DNA-NPR column. The results indicated that the separation of DNA fragments on the anion-exchange HPLC was governed not only by size, but also by nucleotide sequence: even DNA fragments with the same size and the same base content could be separated on this column. Taking advantage of this characteristic feature of the anion-exchange HPLC, we could readily fractionate human cDNAs with practically acceptable recovery and high resolution. Furthermore, the combination of HPLC and gel electrophoresis realized separation of a mixture of DNA fragments in a two-dimensional pattern.
Yamakawa
H
H
Kazusa DNA Research Institute, 1532-3 Yana-Uchino, Chiba, Kisarazu, 292, Japan.
Higashino
K i
Ki
Ohara
O
O
D001483:Base Sequence; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004247:DNA / Q000032:analysis; D004262:DNA Restriction Enzymes; D018076:DNA, Complementary / Q000032:analysis* / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008969:Molecular Sequence Data
D018076:DNA, Complementary; D004247:DNA; D004262:DNA Restriction Enzymes