Polymerase chain reaction-capillary electrophoresis genetic analysis microdevice with in-line affinity capture sample injection.
Polymerase chain reaction-capillary electrophoresis genetic analysis microdevice with in-line affinity capture sample injection.
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DOI：
10.1021/ac802057f
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发表时间：
2009-01
影响因子：
7.4
通讯作者：
Numrin Thaitrong;N. Toriello;Nadia Del Bueno;R. Mathies
中科院分区：
文献类型：
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作者：
Numrin Thaitrong;N. Toriello;Nadia Del Bueno;R. Mathies
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.
