Multiplex Single-Molecule DNA Barcoding Using an Oligonucleotide Ligation Assay.

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 J

journal_title

Biophysical journal

authors

Severins I,Szczepaniak M,Joo C

doi

10.1016/j.bpj.2018.08.013

subject

Has Abstract

pub_date

2018-09-18 00:00:00

pages

957-967

issue

6

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(18)30965-2

journal_volume

115

pub_type

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