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
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: | https://repository.cshl.edu/id/eprint/25244 |
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