Salvaging recombinants from low-efficiency ligase reactions for more efficient subcloning
Certain types of ligase reactions can be problematic, such as those involving PCR products, blunt-ends and multiple DNA inserts. A simple PCR-based strategy was developed to overcome cloning difficulties with these inefficient ligase reactions. After an initial ligase reaction, primers complementary...
| Veröffentlicht in: | BioTechniques. - 1988. - 18(1995), 4 vom: 15. Apr., Seite 644-6, 648, 650 |
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| Format: | Aufsatz |
| Sprache: | English |
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1995
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| Zugriff auf das übergeordnete Werk: | BioTechniques |
| Schlagworte: | Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Technical Report DNA Primers DNA, Recombinant Receptors, Erythropoietin Ligases EC 6.- |
| Zusammenfassung: | Certain types of ligase reactions can be problematic, such as those involving PCR products, blunt-ends and multiple DNA inserts. A simple PCR-based strategy was developed to overcome cloning difficulties with these inefficient ligase reactions. After an initial ligase reaction, primers complementary to the vector are utilized to amplify the DNA fragment from (the few) successful recombinants in the ligation mixture. This DNA fragment is processed for use in a more conventional and straightforward ligase reaction. We demonstrate the potential of the technique by applying it to a variety of difficult ligase reactions |
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| Beschreibung: | Date Completed 09.08.1995 Date Revised 14.11.2007 published: Print Citation Status MEDLINE |
| ISSN: | 1940-9818 |