Membrane-based hybridization capture of intracellular peptide nucleic acid.
Peptide nucleic acid (PNA) is a chimeric oligonucleotide with nucleotide-derived bases and a peptide backbone. Compared with natural nucleotides, PNA has several advantages, including improved stability and high sequence discrimination during duplex formation. Despite its potential for therapeutic application, analysis technologies have not been generalized, mainly due to ambiguous physiochemical properties resembling those of nucleic acids as well as protein. Here we present a PNA detection method: sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by electrotransfer to a Western blotting membrane and then hybridization with a radiolabeled oligonucleotide probe. This method is useful for evaluating the quality of synthetic PNA and determining its intracellular localization.
Shin
Duckhyang
D
Virus Research Laboratory, Mogam Biotechnology Research Institute, Bojeong-dong, Yongin, Gyeonggi-do 449-913, South Korea.
Nam
Minwoo
M
Yoon
Yeup
Y
Kim
Meehyein
M
D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D035683:MicroRNAs / Q000737:chemistry; D009693:Nucleic Acid Hybridization / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry; D020135:Peptide Nucleic Acids / Q000737:chemistry* / Q000378:metabolism
C531907:MIRN122 microRNA, human; D035683:MicroRNAs; D015345:Oligonucleotide Probes; D020135:Peptide Nucleic Acids