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Parallel synthesis of DNA in multipl...

Blair, Sarah.

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Parallel synthesis of DNA in multiplexed capillaries.

Record Type:	Language materials, printed : Monograph/item
Title/Author:	Parallel synthesis of DNA in multiplexed capillaries./
Author:	Blair, Sarah.
Description:	203 p.
Notes:	Adviser: Franco Cerrina.
Contained By:	Dissertation Abstracts International68-12B.
Subject:	Biology, Bioinformatics. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3294070
ISBN:	9780549385639

Parallel synthesis of DNA in multiplexed capillaries.
Blair, Sarah.

Parallel synthesis of DNA in multiplexed capillaries. - 203 p.

Adviser: Franco Cerrina.

Thesis (Ph.D.)--The University of Wisconsin - Madison, 2007.

The ability to produce synthetic genes has become increasingly important in the emerging field of synthetic biology. The chemical synthesis of single stranded DNA fragments (oligonucleotides) and their subsequent assembly into functioning genes through base pairing, was first achieved in the 1960. Since then, the technology has been refined and automated and today short (¡100 base) oligonucleotides are readily available. However, the demand for kilobase-pair genes, which require thousands of construction oligomers, has lead to a need for a higher-throughput, parallel synthesis technology. Using DNA microarrays to produce large numbers of sequences (700,000) is being explored. But using microarrays means a high cost-per-sequence for small pools of oligos (1-1000) and low sequence quantity from complex arrays (10,000-1,000,000 sequences). This, along with accessibility issues for the average lab, has left a gap in mid-range (500-5000 sequences) parallel olignucleotide production. In order to fill this need, I have developed a method for synthesizing short oligonucleotides in a glass capillary. This new technique uses photo-labile 3-nitrophenylpropyloxycarbonyl (NPPOC) chemistry with UV-LEDs as discreet illumination sources. Custom platforms for fluid delivery, LED control, and synthesis support were designed and engineered for this method. Oligonucleotides were synthesized in single capillaries and in two capillaries run in parallel. Using these building blocks, I was able to assemble 100 and 260 base-pair genes. Future refinement and scaling of this system will lead to a simple, reliable method for gene synthesis.

ISBN: 9780549385639Subjects--Topical Terms:

1018415
Biology, Bioinformatics.

Parallel synthesis of DNA in multiplexed capillaries.
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020 $a 9780549385639
035 $a (UMI)AAI3294070
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100 1 $a Blair, Sarah. $3 1272244
245 1 0 $a Parallel synthesis of DNA in multiplexed capillaries.
300 $a 203 p.
500 $a Adviser: Franco Cerrina.
500 $a Source: Dissertation Abstracts International, Volume: 68-12, Section: B, page: 7777.
502 $a Thesis (Ph.D.)--The University of Wisconsin - Madison, 2007.
520 $a The ability to produce synthetic genes has become increasingly important in the emerging field of synthetic biology. The chemical synthesis of single stranded DNA fragments (oligonucleotides) and their subsequent assembly into functioning genes through base pairing, was first achieved in the 1960. Since then, the technology has been refined and automated and today short (¡100 base) oligonucleotides are readily available. However, the demand for kilobase-pair genes, which require thousands of construction oligomers, has lead to a need for a higher-throughput, parallel synthesis technology. Using DNA microarrays to produce large numbers of sequences (700,000) is being explored. But using microarrays means a high cost-per-sequence for small pools of oligos (1-1000) and low sequence quantity from complex arrays (10,000-1,000,000 sequences). This, along with accessibility issues for the average lab, has left a gap in mid-range (500-5000 sequences) parallel olignucleotide production. In order to fill this need, I have developed a method for synthesizing short oligonucleotides in a glass capillary. This new technique uses photo-labile 3-nitrophenylpropyloxycarbonyl (NPPOC) chemistry with UV-LEDs as discreet illumination sources. Custom platforms for fluid delivery, LED control, and synthesis support were designed and engineered for this method. Oligonucleotides were synthesized in single capillaries and in two capillaries run in parallel. Using these building blocks, I was able to assemble 100 and 260 base-pair genes. Future refinement and scaling of this system will lead to a simple, reliable method for gene synthesis.
590 $a School code: 0262.
650 4 $a Biology, Bioinformatics. $3 1018415
650 4 $a Biology, Genetics. $3 1017730
650 4 $a Engineering, Electronics and Electrical. $3 626636
690 $a 0369
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690 $a 0715
710 2 $a The University of Wisconsin - Madison. $3 626640
773 0 $t Dissertation Abstracts International $g 68-12B.
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790 1 0 $a Cerrina, Franco, $e advisor
791 $a Ph.D.
792 $a 2007
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