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SOP_DNA_QIAFastDNA_DTU_v3.pdf (412.56 kB)

SOP - DNA Isolation QIAamp Fast DNA Stool Modified

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Version 4 2016-10-18, 16:26
Version 3 2016-10-18, 16:04
Version 2 2016-07-19, 06:55
Version 1 2016-07-09, 11:40
journal contribution
posted on 2016-10-18, 16:26 authored by Berith E. Knudsen, Lasse Bergmark, Sünje Johanna PampSünje Johanna Pamp
This protocol for DNA isolation is based on the Qiagen kit QIAamp Fast DNA stool mini kit (cat. no. 51604). The method has been tested using different sample matrices, e.g. human and animal feces and sewage. The protocol “Isolation of DNA from Stool for Pathogen Detection” supplied with the kit has been modified in order to increase the DNA yield from these complex samples while maintaining high quality of the DNA. The main modification to the protocol supplied with the kit is the addition of a bead-beating step in the beginning of the isolation to facilitate increasing cell lysis, e.g. of Gram-positive bacteria. Additional modifications have been included as described herein. For example, for the extraction of DNA from sewage specimens a few additional steps are included to further increase the DNA yield.

If you use this method, please cite:

Berith E. Knudsen, Lasse Bergmark, Patrick Munk, Oksana Lukjancenko, Anders Priemé, Frank M. Aarestrup, Sünje J. Pamp (2016) Impact of Sample Type and DNA Isolation Procedure on Genomic Inference of Microbiome Composition. mSystems Oct 2016, 1 (5) e00095-16; DOI: 10.1128/mSystems.00095-16

http://msystems.asm.org/content/1/5/e00095-16




Funding

FP7 (613754), Horizon2020 (643476), Villum Fonden (VWR023052), Carlsberfondet (2013_01_0377)

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