Efficient molecular cloning of environmental DNA from geothermal sediments

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Date

2002

Journal Title

Journal ISSN

Volume Title

Publisher

Kluwer Academic Publishers

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.

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Keywords

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.