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Microfabricated integrated DNA analy...

Woolley, Adam Thomas.

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Microfabricated integrated DNA analysis systems.

紀錄類型:	書目-語言資料,印刷品 : Monograph/item
正題名/作者:	Microfabricated integrated DNA analysis systems./
作者:	Woolley, Adam Thomas.
面頁冊數:	140 p.
附註:	Chair: Richard A. Mathies.
Contained By:	Dissertation Abstracts International59-03B.
標題:	Chemistry, Analytical. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9827157
ISBN:	0591794926

Microfabricated integrated DNA analysis systems.
Woolley, Adam Thomas.

Microfabricated integrated DNA analysis systems. - 140 p.

Chair: Richard A. Mathies.

Thesis (Ph.D.)--University of California, Berkeley, 1997.

Microfabricated electrophoretic DNA analysis systems are developed and utilized for a variety of nucleic acid determinations. Capillary electrophoresis (CE) chips are produced by first chemically etching $\rm {\approx}8\ \mu m$-deep by 30-150 $\rm \mu m$-wide channels in a photolithographically masked

ISBN: 0591794926Subjects--Topical Terms:

586156
Chemistry, Analytical.

Microfabricated integrated DNA analysis systems.
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245 1 0 $a Microfabricated integrated DNA analysis systems.
300 $a 140 p.
500 $a Chair: Richard A. Mathies.
500 $a Source: Dissertation Abstracts International, Volume: 59-03, Section: B, page: 1090.
502 $a Thesis (Ph.D.)--University of California, Berkeley, 1997.
520 $a Microfabricated electrophoretic DNA analysis systems are developed and utilized for a variety of nucleic acid determinations. Capillary electrophoresis (CE) chips are produced by first chemically etching $\rm {\approx}8\ \mu m$-deep by 30-150 $\rm \mu m$-wide channels in a photolithographically masked $5 \times 7.5$ cm glass substrate, followed by thermally bonding the etched piece to a second substrate to create a glass sandwich structure. Reproducible, high-resolution separations of dsDNA restriction fragments and PCR products are demonstrated in ${<}2$ minutes using a replaceable sieving matrix (0.75% hydroxyethylcellulose, 40 mM TAE, pH 8.2) that permits repeated filling and reuse of the same microchannel. With a higher-resolution denaturing polyacrylamide gel these same CE chips provide short tandem repeat genotyping in ${\approx}5$ minutes and DNA sequencing separations in ${\approx}10$ minutes. The resolution is sufficient $({>}0.5)$ for four-color DNA sequencing to ${\approx}200$ bases, and methods are discussed for extending the read length to ${>}400$ bases. A microfabricated silicon thermal cycler attached as the injection reservoir of a glass CE chip is used for on-chip sample preparation. Direct integration of PCR and CE on the same microdevice provides rapid amplification $({\approx}15$ minutes) and analysis $({\approx}120$ seconds) of DNA samples, while eliminating manual reagent transfer steps. These integrated PCR-CE microdevices are utilized to identify diagnostic targets in genomic Salmonella DNA and to measure PCR amplification in real time. Additionally, microfabricated capillary array electrophoresis chips are developed and used to size 12 different DNA samples in ${<}160$ seconds, using either one-or two-color labeling with intercalation dyes and laser-excited confocal-fluorescence scanning detection. These microdevices are used for high-throughput genotyping of HLA-H, a candidate gene for the diagnosis of hereditary hemochromatosis. Finally, microfabricated CE chips with integrated electrochemical detection provide sensitive analysis with minimal interference from the electrophoresis field, through photolithographic placement of the working electrode just outside the separation channel. Separations of neurotransmitters in ${<}100$ seconds and sizing of DNA restriction fragments and PCR products demonstrate the utility of these electrochemical chips. These integrated microdevices are bringing to reality the "laboratory-on-a-chip" concept.
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650 4 $a Chemistry, Biochemistry. $3 1017722
650 4 $a Chemistry, Physical. $3 560527
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710 2 0 $a University of California, Berkeley. $3 687832
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791 $a Ph.D.
792 $a 1997
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