Wilkinson, Dianne E.Jaenicke, ThomasCowan, Donald A.2010-10-212010-10-212002Wilkinson, D., Jaenicke, T., Cowan, D.A. (2002). Efficient cloning of DNA from environmental sources. Biotechnology Letters, 24(2):155-161.http://hdl.handle.net/10566/161An 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.enThis 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.Culture-independent cloningEnvironmental DNA libraryGeothermal sedimentMetagenomeArticle