東華大學圖書館 |

Language: English

Help

回圖書館首頁

手機版館藏查詢

Back

Switch To: Labeled | MARC Mode | ISBD

Microfluidic techniques for DNA melt...

Sundberg, Scott Owen.

Linked to FindBook

Google Book

Amazon

博客來

Microfluidic techniques for DNA melting analysis and digital polymerase chain reaction.

Record Type:	Language materials, printed : Monograph/item
Title/Author:	Microfluidic techniques for DNA melting analysis and digital polymerase chain reaction./
Author:	Sundberg, Scott Owen.
Description:	101 p.
Notes:	Source: Dissertation Abstracts International, Volume: 72-02, Section: B, page: 6860.
Contained By:	Dissertation Abstracts International72-02B.
Subject:	Biology, Molecular. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3432489
ISBN:	9781124369914

Microfluidic techniques for DNA melting analysis and digital polymerase chain reaction.
Sundberg, Scott Owen.

Microfluidic techniques for DNA melting analysis and digital polymerase chain reaction. - 101 p.

Source: Dissertation Abstracts International, Volume: 72-02, Section: B, page: 6860.

Thesis (Ph.D.)--The University of Utah, 2010.

Microfluidic methods were applied to nucleic acid mutation identification and quantification. DNA melting analysis interrogation volumes were reduced 4 orders of magnitude (down to 1 nL volumes) from commercial instrumentation, allowing less reagent consumption while yielding adequate signal for genotyping and scanning of polymerase chain reaction (PCR) products. A microfluidic instrument was developed for digital PCR applications, using a spinning plastic disk patterned by xurography. The platform offers faster thermocycling times (30 cycles in &sim;12 min), simplified fluid partitioning, and a less expensive disposable when compared to currently available digital PCR platforms. PCR within the disk was validated by quantifying plasmid DNA sample using "on/off" fluorescence interrogation across 1000 wells (30 nL/well) at varying template concentration. A 94% PCR efficiency and product amplification specificity were determined by aggregate real-time PCR and melting analysis. The technique of quasi-digital PCR was also applied within this platform, wherein a single mutation copy was preferentially amplified from a large background of wild-type DNA, to detect and quantify low levels of rare mutations. This method demonstrated a sensitivity of 0.01% (detecting a mutant to wild-type DNA ratio of 43:450000), by mixing known concentrations of an oncogene mutation with thousands of wild-type template copies. Statistic analysis tools were constructed in order to interpret digital PCR data, with results comparing well to DNA absorption measurements.

ISBN: 9781124369914Subjects--Topical Terms:

1017719
Biology, Molecular.

Microfluidic techniques for DNA melting analysis and digital polymerase chain reaction.
LDR:02452nam a2200277 4500 001 1958383
005 20140412123520.5
008 150210s2010 ||||||||||||||||| ||eng d
020 $a 9781124369914
035 $a (MiAaPQ)AAI3432489
035 $a AAI3432489
040 $a MiAaPQ $c MiAaPQ
100 1 $a Sundberg, Scott Owen. $3 2093452
245 1 0 $a Microfluidic techniques for DNA melting analysis and digital polymerase chain reaction.
300 $a 101 p.
500 $a Source: Dissertation Abstracts International, Volume: 72-02, Section: B, page: 6860.
500 $a Adviser: Bruce K. Gale.
502 $a Thesis (Ph.D.)--The University of Utah, 2010.
520 $a Microfluidic methods were applied to nucleic acid mutation identification and quantification. DNA melting analysis interrogation volumes were reduced 4 orders of magnitude (down to 1 nL volumes) from commercial instrumentation, allowing less reagent consumption while yielding adequate signal for genotyping and scanning of polymerase chain reaction (PCR) products. A microfluidic instrument was developed for digital PCR applications, using a spinning plastic disk patterned by xurography. The platform offers faster thermocycling times (30 cycles in &sim;12 min), simplified fluid partitioning, and a less expensive disposable when compared to currently available digital PCR platforms. PCR within the disk was validated by quantifying plasmid DNA sample using "on/off" fluorescence interrogation across 1000 wells (30 nL/well) at varying template concentration. A 94% PCR efficiency and product amplification specificity were determined by aggregate real-time PCR and melting analysis. The technique of quasi-digital PCR was also applied within this platform, wherein a single mutation copy was preferentially amplified from a large background of wild-type DNA, to detect and quantify low levels of rare mutations. This method demonstrated a sensitivity of 0.01% (detecting a mutant to wild-type DNA ratio of 43:450000), by mixing known concentrations of an oncogene mutation with thousands of wild-type template copies. Statistic analysis tools were constructed in order to interpret digital PCR data, with results comparing well to DNA absorption measurements.
590 $a School code: 0240.
650 4 $a Biology, Molecular. $3 1017719
650 4 $a Chemistry, Analytical. $3 586156
690 $a 0307
690 $a 0486
710 2 $a The University of Utah. $b Bioengineering. $3 1672369
773 0 $t Dissertation Abstracts International $g 72-02B.
790 $a 0240
791 $a Ph.D.
792 $a 2010
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3432489