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.
Description
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.