机构：[1]Yunnan Institute of Digestive Disease,The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan Province, China昆明医科大学附属第一医院[2]Department of Reproduction and Genetics,The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan Province, China昆明医科大学附属第一医院[3]The First People’s Hospital of Yunnan Province, Kunming, Yunnan Province, China云南省第一人民医院
Formalin fixation and paraffin embedding is widely used for convenient and long-term storage of tumor tissue and precious sources to perform genetic studies. However, DNA fragmentation is one of the major flaws of genomic DNA isolation from formalin fixation tissues, which limits its further usage. Here, we present an improved method for isolating high-quality genomic DNA from formalin fixation tissue. We obtained high-quality genomic DNA of more than 20 kb from samples frozen for more than 2 years. Furthermore, to verify DNA quality, the whole mitochondrial DNA (mtDNA) genomes from the normal and tumor tissue of the same patient were successfully amplified with two overlapping PCR fragments comprising more than 8379 bp in length for each fragment. In addition, the whole genomes were sequenced with a 48-well based primer panel in order to avoid potential sequencing errors from artificial recombination, which was further confirmed with an mtDNA phylogenetic strategy. Our improved DNA extraction method from formalin fixation tissue and sequencing strategy for entire mtDNA genomes will generate unambiguous sequence analysis results for clinical samples.
基金：
Foundation for Science and Technology Planning Project of Yunnan Provincial Bureau of Health [2012WS063]; Yunnan Institute of Digestive Disease Institute [2014NS121, 2014NS122]; China Postdoctoral Science FoundationChina Postdoctoral Science Foundation [2014M562513XB]; National Science Foundation of ChinaNational Natural Science Foundation of China [81360069]; Academician Workstation of Yunnan Province; Foundation for Innovative Group of Gastrointestinal Surgery of Yunnan Province [2012HC013]; Foundation of Medical Leading Talent of Yunnan Province [L-201205]
第一作者机构：[1]Yunnan Institute of Digestive Disease,The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan Province, China[2]Department of Reproduction and Genetics,The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan Province, China
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通讯作者：
推荐引用方式(GB/T 7714)：
Wang H. W.,Xu Y.,Zhang H. F.,et al.Improved protocol for extracting genomic DNA from frozen formalin-fixed tissue resulting in high-quality whole mtDNA[J].GENETICS AND MOLECULAR RESEARCH.2016,15(3):doi:10.4238/gmr.15037972.
APA：
Wang, H. W.,Xu, Y.,Zhang, H. F.,Zeng, Y. J.,Ren, L....&Wang, K. H..(2016).Improved protocol for extracting genomic DNA from frozen formalin-fixed tissue resulting in high-quality whole mtDNA.GENETICS AND MOLECULAR RESEARCH,15,(3)
MLA：
Wang, H. W.,et al."Improved protocol for extracting genomic DNA from frozen formalin-fixed tissue resulting in high-quality whole mtDNA".GENETICS AND MOLECULAR RESEARCH 15..3(2016)
