Blunt-end and single-strand ligations by Escherichia coli ligase: influence on an in vitro amplication scheme

Barringer, K. J., Orgel, L., Wahl, G., Gingeras, T. R. (1990) Blunt-end and single-strand ligations by Escherichia coli ligase: influence on an in vitro amplication scheme. Gene, 89 (1). pp. 117-122. ISSN 0378-1119

URL: http://www.ncbi.nlm.nih.gov/pubmed/2197176
DOI: 10.1016/0378-1119(90)90213-b

Abstract

A ligase-based, in vitro DNA amplication system (LAR) has been described by Wu and Wallace [Genomics 4 (1989) 560–569]. This strategy is based on the ability of a DNA ligase to join the 5′ phosphate of one DNA molecule to the 3′ hydroxyl of a second during a nick-closing reaction. Escherichia coli DNA ligase has been used in place of the T4 DNA ligase in our study in order to limit template-independent ligation activities, which lower the sensitivity of this amplication procedure. The results of this studyes indicate that E. coli ligase also joins blunt-ended DNA molecules and some single-stranded oligodeoxyribonucleotides, in the absence of a complementary template, with an efficiency which is sensitive to both the concentrations of DNA substrate and enzyme.

Item Type: Paper
Uncontrolled Keywords: Recombinant DNA T4 ligase in vitro nucleic acid amplification nick closing blunt-end ligation
Subjects: Investigative techniques and equipment
organism description > bacteria
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene amplification
CSHL Authors:
Communities: CSHL labs > Gingeras lab
Depositing User: CSHL Librarian
Date: 1990
Date Deposited: 14 Mar 2012 14:27
Last Modified: 28 Mar 2014 20:52
Related URLs:
URI: http://repository.cshl.edu/id/eprint/25244

Actions (login required)

Administrator's edit/view item Administrator's edit/view item
CSHL HomeAbout CSHLResearchEducationNews & FeaturesCampus & Public EventsCareersGiving