creators_name: Kumar, Raj
creators_name: Rajagopal, Kammara
type: article
datestamp: 2011-11-30 11:37:38
lastmod: 2011-11-30 11:37:38
metadata_visibility: show
title: Single-step overlap-primer-walk polymerase chain reaction for multiple mutagenesis without overlap extension.
ispublished: pub
subjects: QD
full_text_status: restricted
note: copyright of this article belongs to Elsevier Science
abstract: 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.
date: 2008-06-01
date_type: published
publication: Analytical biochemistry
volume: 377
number: 1
publisher: Elsevier Science
pagerange: 105-107
refereed: TRUE
issn: 1096-0309
official_url: http://linkinghub.elsevier.com/retrieve/pii/S0003-2697(08)00133-4
related_url_url: http://linkinghub.elsevier.com/retrieve/pii/S0003-2697(08)00133-4
related_url_type: pub
citation:   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     
document_url: http://crdd.osdd.net/open/610/1/rajgopal08.pdf