Nucleic Acids Research Advance Access originally published online on June 18, 2007
Nucleic Acids Research 2007 35(12):e85; doi:10.1093/nar/gkm433
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Nucleic Acids Research, 2007, Vol. 35, No. 12 e85
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Novel zinc-based fixative for high quality DNA, RNA and protein analysis
1Department of Histopathology, Division of Investigative Sciences, Imperial College London, Hammersmith Hospital Campus, Ducane Road, London, W12 ONN, UK, 2Department of Chemistry, South Kensington Campus, Imperial College London, London, SW7 2AZ, UK, 3Experimental Pathology Laboratory, Cancer Research UK London Research Institute, Lincoln's Inn Fields Laboratories, WC2A 3PX, London, UK and 4Molecular Pathogenesis Unit, NINDS, National Institutes of Health, Bldg 10, Bethesda, MD 20892, USA
*To whom correspondence should be addressed. Tel: +44-208-3832445; Fax: +44-208-7407417; Email: dimitrios.lykidis00{at}ic.ac.uk
Received February 28, 2007. Revised May 12, 2007. Accepted May 14, 2007.
We have developed a reliable, cost-effective and non-toxic fixative to meet the needs of contemporary molecular pathobiology research, particularly in respect of RNA and DNA integrity. The effects of 25 different fixative recipes on the fixed quality of tissues from C57BL/6 mice were investigated. Results from IHC, PCR, RT–PCR, RNA Agilent Bioanalyser and Real-Time PCR showed that a novel zinc-based fixative (Z7) containing zinc trifluoroacetate, zinc chloride and calcium acetate was significantly better than the standard zinc-based fixative (Z2) and neutral buffered formalin (NBF) for DNA, RNA and protein preservation. DNA sequences up to 2.4 kb in length and RNA fragments up to 361 bp in length were successfully amplified from Z7 fixed tissues, as demonstrated by PCR, RT–PCR and Real-Time PCR. Total protein analysis was achieved using 2-D gel electrophoresis. In addition, nucleic acids and proteins were very stable over a 6–14-month period. This improved, non-toxic and economical tissue fixative could be applied for routine use in pathology laboratories to permit subsequent genomic/proteomic studies.
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