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Single-step overlap-primer-walk polymerase chain reaction for multiple mutagenesis without overlap extension.

DIR@IMTECH: CSIR-Institute of Microbial Technology

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Title Single-step overlap-primer-walk polymerase chain reaction for multiple mutagenesis without overlap extension.
 
Creator Kumar, Raj
Rajagopal, Kammara
 
Subject QD Chemistry
 
Description We present a simple, single-step, single-tube, and rapid method for introducing a series of mutations into cloned DNA. Polymerase chain reaction (PCR)-based mutagenesis methods have become very prevalent due to their simplicity and efficiency for introducing mutations. Our method, overlap-primer-walk PCR, has several advantages over other published methods. It uses two common oligodeoxyribonucleotides and a series of overlapping primers specific for various mutations. Once common flanking primers are selected, two to three mutations require only one additional primer. Therefore, this method is very useful for introduction of multiple mutations in various sites of the target DNA. We illustrate the usefulness of the method by introducing several mutations into the human TNF-alpha encoding gene.
 
Publisher Elsevier Science
 
Date 2008-06-01
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://crdd.osdd.net/open/610/1/rajgopal08.pdf
Kumar, Raj and Rajagopal, Kammara (2008) Single-step overlap-primer-walk polymerase chain reaction for multiple mutagenesis without overlap extension. Analytical biochemistry, 377 (1). pp. 105-107. ISSN 1096-0309
 
Relation http://linkinghub.elsevier.com/retrieve/pii/S0003-2697(08)00133-4
http://crdd.osdd.net/open/610/