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Title: Construction of a microfluidic chip, using dried-down reagents, for LATE-PCR amplification and detection of single-stranded DNA.
Authors: Jia, Yan wei
Mak, Pui In (麥沛然)
Conner, Massey
Martins, Rui Paulo Da Silva
Wangh, Lawrence J.
Issue Date: 7-Dec-2013
Publisher: Royal Society of Chemistry
Citation: Lab on a Chip, Dec 2013, Vol. 13, Issue 23, p. 4635-4641
Abstract: LATE-PCR is an advanced form of non-symmetric PCR that efficiently generates single-stranded DNA which can readily be characterized at the end of amplification by hybridization to low-temperature fluorescent probes. We demonstrate here for the first time that monoplex and duplex LATE-PCR amplification and probe target hybridization can be carried out in double layered PDMS microfluidics chips containing dried reagents. Addition of a set of reagents during dry down overcomes the common problem of single-stranded oligonucleotide binding to PDMS. These proof-of-principle results open the way to construction of inexpensive point-of-care devices that take full advantage of the analytical power of assays built using LATE-PCR and low-temperature probes.
ISSN: 1473-0197
Access: View full-text via DOI

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