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Novel zinc-based fixative for high quality DNA, RNA and protein analysis

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. R...

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Detalles Bibliográficos
Autores principales: Lykidis, Dimitrios, Van Noorden, Susan, Armstrong, Alan, Spencer-Dene, Bradley, Li, Jie, Zhuang, Zhengping, Stamp, Gordon W. H.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1919503/
https://www.ncbi.nlm.nih.gov/pubmed/17576663
http://dx.doi.org/10.1093/nar/gkm433
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author Lykidis, Dimitrios
Van Noorden, Susan
Armstrong, Alan
Spencer-Dene, Bradley
Li, Jie
Zhuang, Zhengping
Stamp, Gordon W. H.
author_facet Lykidis, Dimitrios
Van Noorden, Susan
Armstrong, Alan
Spencer-Dene, Bradley
Li, Jie
Zhuang, Zhengping
Stamp, Gordon W. H.
author_sort Lykidis, Dimitrios
collection PubMed
description 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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spelling pubmed-19195032007-07-24 Novel zinc-based fixative for high quality DNA, RNA and protein analysis Lykidis, Dimitrios Van Noorden, Susan Armstrong, Alan Spencer-Dene, Bradley Li, Jie Zhuang, Zhengping Stamp, Gordon W. H. Nucleic Acids Res Methods Online 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. Oxford University Press 2007-06 2007-06-18 /pmc/articles/PMC1919503/ /pubmed/17576663 http://dx.doi.org/10.1093/nar/gkm433 Text en © 2007 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ 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.
spellingShingle Methods Online
Lykidis, Dimitrios
Van Noorden, Susan
Armstrong, Alan
Spencer-Dene, Bradley
Li, Jie
Zhuang, Zhengping
Stamp, Gordon W. H.
Novel zinc-based fixative for high quality DNA, RNA and protein analysis
title Novel zinc-based fixative for high quality DNA, RNA and protein analysis
title_full Novel zinc-based fixative for high quality DNA, RNA and protein analysis
title_fullStr Novel zinc-based fixative for high quality DNA, RNA and protein analysis
title_full_unstemmed Novel zinc-based fixative for high quality DNA, RNA and protein analysis
title_short Novel zinc-based fixative for high quality DNA, RNA and protein analysis
title_sort novel zinc-based fixative for high quality dna, rna and protein analysis
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1919503/
https://www.ncbi.nlm.nih.gov/pubmed/17576663
http://dx.doi.org/10.1093/nar/gkm433
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