Please use this identifier to cite or link to this item: http://hdl.handle.net/1822/51593

TitleColony PCR
Author(s)Azevedo, Flávio
Pereira, Humberto
Johansson, Björn
KeywordsColony
Direct lysis
Escherichia coli
PCR
Saccharomyces cerevisiae
Yeast
Issue date1-Jan-2017
PublisherSpringer
JournalMethods in Molecular Biology
CitationAzevedo F., Pereira H., Johansson B. (2017) Colony PCR. In: Domingues L. (eds) PCR. Methods in Molecular Biology, vol 1620. Springer, New York, NY
Abstract(s)Escherichia coli and Saccharomyces cerevisiae are currently the two most important organisms in synthetic biology. E.coli is almost always used for fundamental DNA manipulation while yeast is the simplest host system for studying eukaryotic gene expression and performing large scale DNA assembly. Yeast expression studies may also require altering of the chromosomal DNA by homologous recombination. All these studies require the verification of the expected DNA sequence and the fastest method of screening is colony PCR, which is direct PCR of DNA in cells without prior DNA purification. Colony PCR is hampered by the difficulty of releasing DNA into the PCR mix and the presence of PCR inhibitors. We hereby present one protocol for E. coli and two protocols for S. cerevisiae differing in efficiency and complexity as well as an overview of past and possible future developments of efficient S. cerevisiae colony PCR protocols.
TypebookPart
URIhttp://hdl.handle.net/1822/51593
ISBN978-1-4939-7059-9
e-ISBN978-1-4939-7060-5
DOI10.1007/978-1-4939-7060-5_8
ISSN1064-3745
Publisher versionhttps://link.springer.com/protocol/10.1007/978-1-4939-7060-5_8#citeas
Peer-Reviewedyes
AccessclosedAccess
Appears in Collections:DBio - Livros e Capítulos de Livros/Books and Books Chapters

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