Efficient molecular cloning of environmental DNA from geothermal sediments

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 polym...

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Bibliographic Details
Published in:Biotechnology letters 2002-01, Vol.24 (2), p.155-161
Main Authors: WILKINSON, Dianna E, JEANICKE, Thomas, COWAN, Don A
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Fundamental and applied biological sciences. Psychology
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Summary: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.
ISSN:0141-5492
1573-6776
DOI:10.1023/A:1013825600643