Quantification of damage in DNA recovered from highly degraded samples – a case study on DNA in faeces 英文参考文献.docVIP

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Quantification of damage in DNA recovered from highly degraded samples – a case study on DNA in faeces 英文参考文献.doc

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Quantification of damage in DNA recovered from highly degraded samples – a case study on DNA in faeces 英文参考文献

Frontiers in Zoology BioMedCentral Methodology Open Access Quantification of damage in DNA recovered from highly degraded samples – a case study on DNA in faeces Bruce E Deagle*1,2, J Paige Eveson3 and Simon N Jarman2 Address: 1School of Zoology, University of Tasmania, Box 252-05, Hobart, Tasmania, Australia, 2Australian Antarctic Division, Channel Highway, Kingston, Tasmania, Australia and 3CSIRO Marine and Atmospheric Research, Box 1538, Hobart, Tasmania, Australia Email: Bruce E Deagle* - bedeagle@.au; J Paige Eveson - paige.eveson@csiro.au; Simon N Jarman - simon.jarman@.au * Corresponding author Published: 16 August 2006 Received: 13 May 2006 Accepted: 16 August 2006 Frontiers in Zoology 2006, 3:11 doi:10.1186/1742-9994-3-11 This article is available from: /content/3/1/11 ? 2006 Deagle et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: Poorly preserved biological tissues have become an important source of DNA for a wide range of zoological studies. Measuring the quality of DNA obtained from these samples is often desired; however, there are no widely used techniques available for quantifying damage in highly degraded DNA samples. We present a general method that can be used to determine the frequency of polymerase blocking DNA damage in specific gene-regions in such samples. The approach uses quantitative PCR to measure the amount of DNA present at several fragment sizes within a sample. According to a model of random degradation the amount of available template will decline exponentially with increasing fragment size in damaged samples, and the frequency of DNA damage (λ) can be estimated by determining the rate of decline. Results: The method is illustrated th