title: DNA sequence analysis: a general, simple and rapid method for sequencing large oligodeoxyribonucleotide fragments by mapping.
abstract: Several electrophoretic and chromatographic systems have been investigated and compared for sequence analysis of oligodeoxyribonucleotides. Three systems were found to be useful for the separation of a series of sequential degradation products resulting from a labeled oligonucleotide: (I) 2-D electrophoresisdagger; (II) 2-D PEI-cellulose; and (III) 2-D homochromatography. System (III) proved generally most informative regardless of base composition and sequence. Furthermore, only in this system will the omission of an oligonucleotide in a series of oligonucleotides be self-evident from the two-dimensional map. The sequence of up to fifteen nucleotides can be determined solely by the characteristic mobility shifts of its sequential degradation products distributed on the two-dimensional map. With this method, ten nucleotides from the double-stranded region adjacent to the left-hand 3'-terminus and seven from the right-hand 3'-terminus of bacteriophage lambda DNA have been sequenced. Similarly, nine nucleotides from the double-stranded region adjacent to the left-hand 3'-terminus and five nucleotides from the right-hand terminus of bacteriophage phi80 DNA have also been sequenced. The advantages and disadvantages of each separation system with respect to sequence analysis are discussed.
authors.forename: E
authors.initials: E
authors.lastname: Jay
authors.forename: R
authors.initials: R
authors.lastname: Bambara
authors.forename: R
authors.initials: R
authors.lastname: Padmanabhan
authors.forename: R
authors.initials: R
authors.lastname: Wu
mesh_terms: D001483:Base Sequence; D002482:Cellulose / Q000031:analogs & derivatives; D002855:Chromatography, Thin Layer; D004279:DNA, Viral / Q000737:chemistry / Q000302:isolation & purification; D004588:Electrophoresis, Cellulose Acetate; D015180:Electrophoresis, Gel, Two-Dimensional; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000302:isolation & purification; D011094:Polyethyleneimine / Q000031:analogs & derivatives; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; C018367:polyethyleneimine-cellulose; D011094:Polyethyleneimine; D002482:Cellulose