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Facile recovery of individual high-molecular-weight, low-copy-number natural plasmids for genomic sequencing. / Williams, Laura E.; Detter, Chris; Barry, Kerrie; Lapidus, Alla; Summers, Anne O.

In: Applied and Environmental Microbiology, Vol. 72, No. 7, 07.2006, p. 4899-4906.

Research output: Contribution to journal › Article › peer-review

Harvard

Williams, LE, Detter, C, Barry, K, Lapidus, A & Summers, AO 2006, 'Facile recovery of individual high-molecular-weight, low-copy-number natural plasmids for genomic sequencing', Applied and Environmental Microbiology, vol. 72, no. 7, pp. 4899-4906. https://doi.org/10.1128/AEM.00354-06

APA

Williams, L. E., Detter, C., Barry, K., Lapidus, A., & Summers, A. O. (2006). Facile recovery of individual high-molecular-weight, low-copy-number natural plasmids for genomic sequencing. Applied and Environmental Microbiology, 72(7), 4899-4906. https://doi.org/10.1128/AEM.00354-06

Vancouver

Williams LE, Detter C, Barry K, Lapidus A, Summers AO. Facile recovery of individual high-molecular-weight, low-copy-number natural plasmids for genomic sequencing. Applied and Environmental Microbiology. 2006 Jul;72(7):4899-4906. https://doi.org/10.1128/AEM.00354-06

Author

Williams, Laura E. ; Detter, Chris ; Barry, Kerrie ; Lapidus, Alla ; Summers, Anne O. / Facile recovery of individual high-molecular-weight, low-copy-number natural plasmids for genomic sequencing. In: Applied and Environmental Microbiology. 2006 ; Vol. 72, No. 7. pp. 4899-4906.

BibTeX

@article{0ec7191025f043aba67587d517dfd264,
title = "Facile recovery of individual high-molecular-weight, low-copy-number natural plasmids for genomic sequencing",
abstract = "Sequencing of the large (>50 kb), low-copy-number (<5 per cell) plasmids that mediate horizontal gene transfer has been hindered by the difficulty and expense of isolating DNA from individual plasmids of this class. We report here that a kit method previously devised for purification of bacterial artificial chromosomes (BACs) can be adapted for effective preparation of individual plasmids up to 220 kb from wild gram-negative and gram-positive bacteria. Individual plasmid DNA recovered from less than 10 ml of Escherichia coli, Staphylococcus, and Corynebacterium cultures was of sufficient quantity and quality for construction of high-coverage libraries, as shown by sequencing five native plasmids ranging in size from 30 kb to 94 kb. We also report recommendations for vector screening to optimize plasmid sequence assembly, preliminary annotation of novel plasmid genomes, and insights on mobile genetic element biology derived from these sequences. Adaptation of this BAC method for large plasmid isolation removes one major technical hurdle to expanding our knowledge of the natural plasmid gene pool.",
author = "Williams, {Laura E.} and Chris Detter and Kerrie Barry and Alla Lapidus and Summers, {Anne O.}",
year = "2006",
month = jul,
doi = "10.1128/AEM.00354-06",
language = "English",
volume = "72",
pages = "4899--4906",
journal = "Applied Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "7",

}

RIS

TY - JOUR

T1 - Facile recovery of individual high-molecular-weight, low-copy-number natural plasmids for genomic sequencing

AU - Williams, Laura E.

AU - Detter, Chris

AU - Barry, Kerrie

AU - Lapidus, Alla

AU - Summers, Anne O.

PY - 2006/7

Y1 - 2006/7

N2 - Sequencing of the large (>50 kb), low-copy-number (<5 per cell) plasmids that mediate horizontal gene transfer has been hindered by the difficulty and expense of isolating DNA from individual plasmids of this class. We report here that a kit method previously devised for purification of bacterial artificial chromosomes (BACs) can be adapted for effective preparation of individual plasmids up to 220 kb from wild gram-negative and gram-positive bacteria. Individual plasmid DNA recovered from less than 10 ml of Escherichia coli, Staphylococcus, and Corynebacterium cultures was of sufficient quantity and quality for construction of high-coverage libraries, as shown by sequencing five native plasmids ranging in size from 30 kb to 94 kb. We also report recommendations for vector screening to optimize plasmid sequence assembly, preliminary annotation of novel plasmid genomes, and insights on mobile genetic element biology derived from these sequences. Adaptation of this BAC method for large plasmid isolation removes one major technical hurdle to expanding our knowledge of the natural plasmid gene pool.

AB - Sequencing of the large (>50 kb), low-copy-number (<5 per cell) plasmids that mediate horizontal gene transfer has been hindered by the difficulty and expense of isolating DNA from individual plasmids of this class. We report here that a kit method previously devised for purification of bacterial artificial chromosomes (BACs) can be adapted for effective preparation of individual plasmids up to 220 kb from wild gram-negative and gram-positive bacteria. Individual plasmid DNA recovered from less than 10 ml of Escherichia coli, Staphylococcus, and Corynebacterium cultures was of sufficient quantity and quality for construction of high-coverage libraries, as shown by sequencing five native plasmids ranging in size from 30 kb to 94 kb. We also report recommendations for vector screening to optimize plasmid sequence assembly, preliminary annotation of novel plasmid genomes, and insights on mobile genetic element biology derived from these sequences. Adaptation of this BAC method for large plasmid isolation removes one major technical hurdle to expanding our knowledge of the natural plasmid gene pool.

UR - http://www.scopus.com/inward/record.url?scp=33746087277&partnerID=8YFLogxK

U2 - 10.1128/AEM.00354-06

DO - 10.1128/AEM.00354-06

M3 - Article

C2 - 16820486

AN - SCOPUS:33746087277

VL - 72

SP - 4899

EP - 4906

JO - Applied Microbiology

JF - Applied Microbiology

SN - 0099-2240

IS - 7

ER -

ID: 90037965