Oligonucleotide fingerprinting of arrayed genomic DNA sequences using LNA-modified hybridization probes.
Recently, we established a robust method for the detection of hybridization events using a DNA microarray deposited on a nanoporous membrane. Here, in a follow-up study, we demonstrate the performance of this approach on a larger set of LNA-modified oligoprobes and genomic DNA sequences. Twenty-six different LNA-modified 7-mer oligoprobes were hybridized to a set of 66 randomly selected human genomic DNA clones spotted on a nanoporous membrane slide. Subsequently, assay sensitivity analysis was performed using receiver operating characteristic (ROC) curves. Comparison of LNA-modified heptamers and DNA heptamers revealed that the LNA modification clearly improved sensitivity and specificity of hybridization experiment. Clustering analysis was applied in order to test practical performance of hybridization experiments with LNA-modified oligoprobes in recognizing similarity of genomic DNA sequences. Comparing the results with the theoretical sequence clusters, we conclude that the application of LNA-modified oligoprobes allows for reliable clustering of DNA sequences which reflects the underlying sequence homology. Our results show that LNA-modified oligoprobes can be used effectively to unravel sequence similarity of DNA sequences and thus, to characterize the content of unknown DNA libraries.
Liu
Jian-Ping
JP
Department of Vertebrate Genomics, Max Planck Institute for Molecular Genetics, Berlin, Germany.
Drungowski
Mario
M
Nyársik
Lajos
L
Schwartz
Regine
R
Lehrach
Hans
H
Herwig
Ralf
R
Janitz
Michal
M
D016000:Cluster Analysis; D004247:DNA / Q000032:analysis*; D016172:DNA Fingerprinting / Q000379:methods*; D030541:Databases, Genetic; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry*; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense / Q000737:chemistry*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*
D015345:Oligonucleotide Probes; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; C477371:locked nucleic acid; D004247:DNA