Linked Life Data

19343708

Source:http://linkedlifedata.com/resource/pubmed/id/19343708

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
2009-4-3
pubmed:abstractText
This unit describes a simple and efficient DNA engineering method that combines nucleotide sequence alteration, multiple PCR fragment assembly, and directional cloning. PCR primers contain a single deoxyuracil residue (dU), and can be designed to accommodate nucleotide substitutions, insertions, and/or deletions. The primers are then used to amplify DNA in discrete fragments that incorporate a dU at each end. Excision of deoxyuracils results in PCR fragments flanked by unique, overlapping, single-stranded extensions that allow the seamless and directional assembly of customized DNA molecules into a linearized vector. In this way, multi-fragment assemblies, as well as various mutagenic changes, can all be accomplished in a single-format experiment. Two basic protocols on the methods of uracil excision-based engineering are presented, and special attention is given to primer design. The use of a commercially available cloning vector and the preparation of custom vectors are also presented.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
1934-3647
pubmed:author
pubmed:copyrightInfo
Copyright 2009 by John Wiley & Sons, Inc.
pubmed:issnType
Electronic
pubmed:volume
Chapter 3
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
Unit 3.21
pubmed:meshHeading
pubmed:year
2009
pubmed:articleTitle
DNA cloning and engineering by uracil excision.
pubmed:affiliation
New England Biolabs, Ipswich, Massachusetts, USA.
pubmed:publicationType
Journal Article