Cosmid vector design for large scale sequencing: the Pyrococcus furiosus genome

Publication Type dissertation
School or College School of Medicine
Department Human Genetics
Author Yeh, Raymond K.
Title Cosmid vector design for large scale sequencing: the Pyrococcus furiosus genome
Date 2000-05
Description Genomic sequencing can be thought of as a process of disassembling a genome into smaller pieces and subsequent reassembly back into the completed genome. This process involves the representation of the genome in libraries of DNA pieces consisting of fractions of the genome ranging in size from 100s of kilobases to hundreds of bases. In order to facilitate the process of library construction and fractionation of genomes a new cosmid vector was designed. The new cosmid vector features an inducible copy number control. Additional features that facilitate production sequencing were also included. The new vector was used in construction of a genomic library of the archea Pyrococcus furious. This library has an average insert size of 33 kb and has a genomic equivalent of 7X. This library, along with a plasmid library, was used to sequence the entire 1,908,253 bases of the P. furiosus genome. During the sequencing it was observed that stability of the cosmids and subsequent recovery of DNA were inversely proportional to the size of the insert size. Smaller inserts were more stable and had higher recovery yields than larger inserts. Further study determined that large inserts decreased the copy number of the cosmid and also inhibited the ability to induce to higher copy number. Various potential explanations were investigated.
Type Text
Publisher University of Utah
Subject Microbial; Pyrococcus furiosus
Subject MESH Gene Library; Genetics
Dissertation Institution University of Utah
Dissertation Name PhD
Language eng
Relation is Version of Digital reproduction of “Cosmid vector design for large scale sequencing: the Pyrococcus furiosus genome.” Spencer S. Eccles Health Sciences Library. Print version of “Cosmid vector design for large scale sequencing: the Pyrococcus furiosus genome.” available at J. Willard Marriott Library Special Collection. QH9.7 2000 .Y44.
Rights Management © Raymond K. Yeh.
Format Medium application/pdf
Format Extent 1,668,251 bytes
Identifier undthes,5170
Source Original: University of Utah Spencer S. Eccles Health Sciences Library (no longer available).
Master File Extent 1,668,274 bytes
ARK ark:/87278/s6nz89g3
Setname ir_etd
Date Created 2012-04-24
Date Modified 2012-04-24
ID 191755
Reference URL https://collections.lib.utah.edu/details?id=191755
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