SNP typing on the NanoChip electronic microarray.
We describe a single nucleotide polymorphism (SNP) typing protocol developed for the NanoChip electronic microarray. The NanoChip array consists of 100 electrodes covered by a thin hydrogel layer containing streptavidin. An electric currency can be applied to one, several, or all electrodes at the same time according to a loading protocol generated by the user. Biotinylated deoxyribonucleic acid (DNA) is directed to the pad(s) via the electronic field(s) and bound to streptavidin in the hydrogel layer. Subsequently, fluorescently labeled reporter oligos and a stabilizer oligo are hybridized to the bound DNA. Base stacking between the short reporter and the longer stabilizer oligo stabilizes the binding of a matching reporter, whereas the binding of a reporter carrying a mismatch in the SNP position will be relatively weak. Thermal stringency is applied to the NanoChip array according to a reader protocol generated by the user and the fluorescent label on the matching reporter is detected.
Børsting
Claus
C
Department of Forensic Genetics, Institute of Forensic Medicine, University of Copenhagan, Copenhagan, Denmark.
Sanchez
Juan J
JJ
Morling
Niels
N
D001483:Base Sequence; D017931:DNA Primers; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis
D017931:DNA Primers