Efficient molecular cloning of environmental DNA from geothermal sediments
| Title: | Efficient molecular cloning of environmental DNA from geothermal sediments |
| Author: | Wilkinson, Dianne E.; Jaenicke, Thomas; Cowan, Donald A. |
| Inquiries: | dcowan@uwc.ac.za |
| Abstract: | An efficient and simple method for constructing an environmental library using mechanically sheared DNA obtained directly from geothermal sediments is presented. The method is based on blunt-end modification of DNA fragments followed by 3' -adenylation using Vent DNA polymerase and Taq DNA polymerase, respectively. The prepared DNA fragments are then ligated into a TA cloning vector and used in the transformation of Escherichia coli. This method has been successfully applied to the cloning of ORFs derived from uncultivated prokaryotes present in geothermal sediment. |
| Subject: | Culture-independent cloning Environmental DNA library Geothermal sediment Metagenome |
| Citation: | Wilkinson, D., Jaenicke, T., Cowan, D.A. (2002). Efficient cloning of DNA from environmental sources. Biotechnology Letters, 24(2):155-161. DOI 10.1023/A:1013825600643 |
| Rights: | This is the author post-print of an article published by Kluwer Academic Publishers. This version may be freely used in non-commercial applications, subject to full acknowledgement. |
| Type: | Article |
| URI: | http://hdl.handle.net/10566/161 |
| DOI: | http://dx.doi.org/10.1023/A:1013825600643 |
| Date: | 2002 |
| Peer reviewed: | Yes |
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