The separation of oligodeoxynucleotides having a single-base difference by affinity capillary electrophoresis using oligodeoxynucleotide- polyacrylamide conjugate.
We have developed an affinity capillary electrophoresis for gene mutation assay. We show a method for separating a mixture of oligodeoxynucleotides having single-base difference using oligodeoxynucleotide-polyacrylamide conjugate. Because the conjugate behaved similarly to nonionic polyacrylamide in terms of migration rate, it can be used as a pseudoimmobilized affinity ligand in a polyacrylamide-coated capillary. Oligodeoxynucleotide having an oncogene sequence and a version with one base substituted were completely separated by this method. The magnesium-ion concentration was found to be a key factor in achieving efficient separation of oligodeoxynucleotides with the same chain length.
Anada
Takahisa
T
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka, Japan.
Arisawa
Takumi
T
Ozaki
Yoshihisa
Y
Takarada
Tohru
T
Katayama
Yoshiki
Y
Maeda
Mizuo
M
D000180:Acrylic Resins / Q000737:chemistry; D000345:Affinity Labels / Q000737:chemistry*; D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000295:instrumentation / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D011905:Genes, ras / Q000235:genetics; D008274:Magnesium / Q000494:pharmacology; D009691:Nucleic Acid Denaturation / Q000187:drug effects; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification*; D017354:Point Mutation; D013696:Temperature; D000099166:Polyacrylamides
D000180:Acrylic Resins; D000345:Affinity Labels; D008274:Magnesium; D009838:Oligodeoxyribonucleotides; D000099166:Polyacrylamides