Abstract
Plasmid extraction is typically performed to produce template DNA for a desired molecular biological reaction, or set of reactions, such as restriction endonuclease digestion (see Chapter 20), DNA sequencing (see Chapter 22), in vitro mutagenesis (see Chapters 23–26), transformation (see Chapters 4 and 5), transfection, or probe generation. The principal methodologies of extraction, involving either alkaline lysis or boiling are well established (1) and are described in detail in Chapters 8 and 9. These methods are relatively crude but are both inexpensive and rapid, and they yield DNA that is suitable for many molecular biology protocols. DNA of higher purity can be prepared using silica resin-based anion exchange as described in Chapter 10 or by separation on cesium chloride gradients (for details, see ref. 1, Chapter 1, Protocol 10). Plasmid minipreps are a ubiquitous part of molecular biology. They are simple and easy to perform but can be very tedious when large numbers of samples are involved. In recent years, endeavors such as the human genome project have led to the development of methods for the manageable high-throughput extraction of plasmids (2–15).
This is a preview of subscription content, log in via an institution to check access.
References
Sambrook, J. and Russell, D. W. (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Sanger protocols. http://www.sanger.ac.uk/Teams/Team51/. Accessed February 13, 2003.
Gibson, T. J. and Sulston, J. E. (1987) Preparation of large numbers of plasmid DNA samples in microtiter plates by the alkaline lysis method. Gene Anal. Tech. 4, 41–44.
Bankier, T. (1993) M13 phage growth and DNA purification using 96 well microtiter trays. Methods Mol. Biol. 23, 41–45.
Elkin, C. J., Richardson, P. M., Fourcade, H. M., et al. (2001) High-throughput plasmid purification for capillary sequencing. Genome Res. 11, 1269–1274.
Engelstein, M., Aldredge, T. J., Madan, D., et al. (1998) An efficient, automatable template preparation for high throughput sequencing. Microb. Comp. Genomics 3, 237–241.
Garner, H. R., Armstrong, B., and Kramarsky, D. A. (1992) High-throughput DNA preparation system. Genet. Anal. Tech. Appl. 9, 134–139.
Hilbert, H., Lauber, J., Lubenow, H., et al. (2000) Automated sample-preparation technologies in genome sequencing projects. DNA Sequence 11, 193–197.
Itoh, M., Kitsunai, T., Akiyama, J., et al. (1999) Automated filtration-based high-throughput plasmid preparation system. Genome Res. 9, 463–470.
Itoh, M., Carninci, P., Nagaoka, S., et al. (1997) Simple and rapid preparation of plasmid template by a filtration method using microtiter filter plates. Nucleic Acids Res. 25, 1315–1316.
Konecki, D. S. and Phillips, J. J. (1998) TurboPrep II: an inexpensive, high-throughput plasmid template preparation protocol. Biotechniques 24, 286–293.
Marra, M. A., Kucaba, T. A., Hillier, L. W., et al. (1999) High-throughput plasmid DNA purification for 3 cents per sample. Nucleic Acids Res. 27, e37; http://www.nar.oupjournals.org/methods.
Neudecker, F. and Grimm, S. (2000) High-throughput method for isolating plasmid DNA with reduced lipopolysaccharide content. Biotechniques 28, 107–109.
Skowronski, E. W., Armstrong, N., Andersen, G., et al. (2000) Magnetic, microplate-format plasmid isolation protocol for high-yield, sequencing-grade DNA. Biotechniques 29, 786–792.
Kelley, J. M., Field, C. E., Craven, M. B., et al. (1999) High throughput direct end sequencing of BAC clones. Nucleic Acids Res. 27, 1539–1546.
Eppendorf AG, http://www.eppendorf.com. Accessed February 13, 2003.
Invitrogen life-technologies, http://www.lifetechnologies.com. Accessed February 13, 2003.
Macherey-Nagel GmbH, Germany, http://www.mn-net.com. Accessed February 13, 2003.
Promega Corporation, http://www.promega.com/nap/. Accessed February 13, 2003.
Qiagen GmbH, http://www.qiagen.com. Accessed February 13, 2003.
Stratagene, http://www.stratagene.com. Accessed February 13, 2003.
Seradyn, http://www.seradyn.com/. Accessed February 13, 2003.
MWG Biotech, http://www.mwgbiotech.com/html/index.shtml. Accessed February 13, 2003.
Beckman Coulter Inc., http://www.beckman.com/products/pr2.asp. Accessed February 13, 2003.
Genetix Ltd., http://www.genetix.co.uk/. Accessed February 13, 2003.
Quail, M. A. (2002) DNA cloning, in Encyclopedia of the Human Genome, (Cooper, D. N., ed.) Nature, London.
Finnpipette, http://www.finnpipette.com/. Accessed February 13, 2003.
Anachem Ltd., Luton, UK. http://www.anachemlifescience.com.uk. Accessed February 13, 2003.
Matrix Technologies, http://www.apogentdiscoveries.com/. Accessed February 13, 2003.
Tomtec, Connecticut, http://www.tomtec.com/. Accessed February 13, 2003.
Mardis, E. (1994) High-thoughput detergent extraction of M13 subclones for fluorescent DNA sequencing. Nucleic Acids Res. 22, 2173–2175.
Washington University Genome Sequencing Center, http://genome.wustl.edu/tools/protocols/. Accessed February 13, 2003.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc.
About this protocol
Cite this protocol
Quail, M.A. (2003). High-Throughput Plasmid Extraction Using Microtiter Plates. In: Casali, N., Preston, A. (eds) E. coli Plasmid Vectors. Methods in Molecular Biology™, vol 235. Humana Press. https://doi.org/10.1385/1-59259-409-3:89
Download citation
DOI: https://doi.org/10.1385/1-59259-409-3:89
Publisher Name: Humana Press
Print ISBN: 978-1-58829-151-6
Online ISBN: 978-1-59259-409-2
eBook Packages: Springer Protocols