Polymerase chain reaction-capillary electrophoresis genetic analysis microdevice with in-line affinity capture sample injection

Analytical Chemistry
Numrin ThaitrongR A Mathies

Abstract

An integrated polymerase chain reaction (PCR)-capillary electrophoresis (CE) microdevice with an efficient in-line affinity-based injector has been developed for genetic analysis. Double stranded DNA PCR amplicons generated in an integrated 250 nL PCR reactor are captured, purified, and preconcentrated by an oligonucleotide probe immobilized in an in situ polymerized gel matrix followed by thermal release and injection into the CE-separation channel. This in-column injector employs a photopolymerized oligonucleotide-modified acrylamide capture gel to eliminate band broadening and increase the injection efficiency to 100%. The on-chip generated PCR amplicons processed on this microdevice exhibit a 3-5 fold increase in signal intensities and improved resolution compared to our previous T-shaped injector. Multiplex analysis of 191-bp amplicons from Escherichia coli O157 and 256-bp amplicons from E. coli K12 is achieved with a 6-fold increase in resolution. These advances are exploited to successfully detect E. coli O157 in a 500-fold higher background of E. coli K12. This microdevice with in-line affinity capture gel injection provides an improved platform for low-volume, high sensitivity, fully integrated genetic analysis.

References

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Citations

Dec 1, 2010·Analytical Chemistry·Numrin ThaitrongRichard A Mathies
Mar 3, 2010·Analytical Chemistry·Yong ZengRichard A Mathies
Feb 11, 2012·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Fumihiko KitagawaKoji Otsuka
Jul 21, 2011·Se pu = Chinese journal of chromatography·Haifang LiJin-Ming Lin
Aug 28, 2009·Trends in Biotechnology·Peng Liu, Richard A Mathies
Apr 1, 2016·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·David P DuberowRichard A Mathies
May 17, 2012·Electrophoresis·Robert J Meagher, Numrin Thaitrong
Aug 23, 2011·Journal of Micromechanics and Microengineering : Structures, Devices, and Systems·Augusto M Tentori, Amy E Herr
Apr 17, 2010·Chemical Reviews·Chunsun Zhang, Da Xing
Dec 8, 2011·Analytical Chemistry·Avraham BrombergRichard A Mathies

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