AFFINITY GEL-ELECTROPHORESIS OF NUCLEIC-ACIDS - SPECIFIC BASE-SELECTIVE AND SHAPE-SELECTIVE SEPARATION OF DNA AND RNA ON POLYACRYLAMIDE-NUCLEOBASE CONJUGATED GEL
E. Yashima et al., AFFINITY GEL-ELECTROPHORESIS OF NUCLEIC-ACIDS - SPECIFIC BASE-SELECTIVE AND SHAPE-SELECTIVE SEPARATION OF DNA AND RNA ON POLYACRYLAMIDE-NUCLEOBASE CONJUGATED GEL, Journal of chromatography, 654(1), 1993, pp. 159-166
Two types of affinity gels consisting of cross-linked polyacrylamide a
nd affinity ligands possessing nucleic acid bases were prepared. One t
ype of gel was polyacrylamide-poly(vinylnucleobase) conjugated gel, wh
ere the poly(vinylnucleobase) such as poly(9-vinyladenine) (PVAd) bear
ing a nucleobase in the side-chain was entrapped in the gel matrix. Th
e other type of gel, in which a nucleobase such as adenine is chemical
ly bonded to polyacrylamide gel, was prepared by copolymerization of a
crylamide, cross-linker and 9-Vinyladenine. These affinity gels, espec
ially the former, demonstrated characteristic nucleobase- and shape se
lective separation of nucleic acids. The gels showed high affinity for
single-stranded DNA and both single- and double-stranded polynucleoti
des and could separate a double-stranded DNA in mixtures of double-str
anded DNA and polynucleotides. The electrophoretic mobilities of poly(
uridylic acid) and poly(inosinic acid) were selectively retarded even
in the presence of 7 M urea. The electrophoretic behaviours of nucleic
acids on the polyacrylamide-PVAd conjugated gels were compared with t
hose on the agarose-PVAd conjugated gel. The effects of urea, temperat
ure and concentration of PVAd were also examined. The polyacrylamide-P
VAd conjugated gel served to elucidate interactions between PVAd and n
ucleic acids that could not be detected by usual spectroscopic methods
.