Abstract:
:Detection of specific nucleic acid sequences is invaluable in biological studies such as genetic disease diagnostics and genome profiling. Here, we developed a highly sensitive and specific detection method that combines an advanced oligonucleotide ligation assay with multicolor single-molecule fluorescence. We demonstrated that under our experimental conditions, 7-nucleotide long DNA barcodes have the optimal short length to ascertain specificity while being long enough for sufficient ligation. Using four spectrally separated fluorophores to label DNA barcodes, we simultaneously distinguished four DNA target sequences differing by only a single nucleotide. Our single-molecule approach will allow for accurate identification of low-abundance molecules without the need for target DNA preamplification.
journal_name
Biophys Jjournal_title
Biophysical journalauthors
Severins I,Szczepaniak M,Joo Cdoi
10.1016/j.bpj.2018.08.013subject
Has Abstractpub_date
2018-09-18 00:00:00pages
957-967issue
6eissn
0006-3495issn
1542-0086pii
S0006-3495(18)30965-2journal_volume
115pub_type
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