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Single-molecule detection and DNA se...

Kartalov, Emil P.

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Single-molecule detection and DNA sequencing-by-synthesis.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Single-molecule detection and DNA sequencing-by-synthesis./
作者:	Kartalov, Emil P.
面頁冊數:	160 p.
附註:	Source: Dissertation Abstracts International, Volume: 65-09, Section: B, page: 4461.
Contained By:	Dissertation Abstracts International65-09B.
標題:	Biophysics, General. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3147958
ISBN:	0496070592

Single-molecule detection and DNA sequencing-by-synthesis.
Kartalov, Emil P.

Single-molecule detection and DNA sequencing-by-synthesis. - 160 p.

Source: Dissertation Abstracts International, Volume: 65-09, Section: B, page: 4461.

Thesis (Ph.D.)--California Institute of Technology, 2004.

Fluorescence detection has established itself as one of the main techniques of interrogation of biological systems. Extending those techniques to decrease the sample size to single molecules provides an absolute standard for bulk sample calibrations, as well as better insights since individual behavior is observed instead of population averages. We observed a number of fluorophores, including GFP, at the single-molecule level at room temperature. Calibrations gave a correct estimate of bulk surface densities over four orders of magnitude, through an optical, non-invasive, non-destructive means. Novel surface chemistry enabled visualization of single tagged nucleotide incorporations inside DNA immobilized on a glass surface at the single-molecule level. This technology was later extended to successful single-molecule DNA sequencing.

ISBN: 0496070592Subjects--Topical Terms:

1019105
Biophysics, General.

Single-molecule detection and DNA sequencing-by-synthesis.
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502 $a Thesis (Ph.D.)--California Institute of Technology, 2004.
520 $a Fluorescence detection has established itself as one of the main techniques of interrogation of biological systems. Extending those techniques to decrease the sample size to single molecules provides an absolute standard for bulk sample calibrations, as well as better insights since individual behavior is observed instead of population averages. We observed a number of fluorophores, including GFP, at the single-molecule level at room temperature. Calibrations gave a correct estimate of bulk surface densities over four orders of magnitude, through an optical, non-invasive, non-destructive means. Novel surface chemistry enabled visualization of single tagged nucleotide incorporations inside DNA immobilized on a glass surface at the single-molecule level. This technology was later extended to successful single-molecule DNA sequencing.
520 $a At the same time, PDMS microfluidics was developed to provide the plumbing control, speed, and economy of scale for a broad range of applications. Novel surface chemistry anchored DNA to the PDMS microchannels, which allowed sequencing-by-synthesis to be conducted in the microfluidic environment using optical techniques. Materials, device, and architecture problems were also solved. Finally, all technology was put together and successful microfluidic bulk-fluorescence DNA sequencing was demonstrated. The same technology is applicable to any DNA studies in microfluidic environments and can eventually be extended to close the circle to single-molecule detection.
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