Optimal preservation of liver biopsy samples for downstream translational applications

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• 作者：Alana R. Sherker (1)
  Vera Cherepanov (1)
  Zahra Alvandi (1)
  Ryan Ramos (1)
  Jordan J. Feld (1) (2)
  
• 关键词：Liver biopsy preservation ; RNAlater ; Allprotect ; RNA integrity
• 刊名：Hepatology International
• 出版年：2013
• 出版时间：June 2013
• 年：2013
• 卷：7
• 期：2
• 页码：758-766
• 全文大小：533KB
• 参考文献：1. Qiagen. RNAlater product handbook. 2006
  2. Qiagen. Allprotect product handbook. 2011
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  5. Imbeaud S, Graudens E, Boulanger V, Barlet X, Zaborski P, Eveno E, Mueller O, Schroeder A, Auffry C. Towards standardization of RNA quality assessment using user-independent classifiers of microcapillary electrophoresis traces. Nucleic Acids Res. 2005;33:e56 CrossRef
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  7. Berg JM, Tymoczko J, Stryer L. Biochemistry. 5th ed.; 2002. p 118-19, 781-08
  
• 作者单位：Alana R. Sherker (1)
  Vera Cherepanov (1)
  Zahra Alvandi (1)
  Ryan Ramos (1)
  Jordan J. Feld (1) (2)
  
  1. University Health Network, McLaughlin-Rotman Centre for Global Health, Toronto General Research Institute, University of Toronto, Toronto, Canada
  2. Toronto Western Hospital Liver Centre, 399 Bathurst Street 6B FP Rm 158, Toronto, ON, M5T 2S8, Canada
  

文摘

Objective: Molecular analysis of liver biopsy samples from patients requires ideal tissue preservation and handling to yield suitable material for laboratory analysis. Biopsy size, tissue handling and preservation method all may affect the quality and quantity of DNA, RNA and protein that can be extracted from liver biopsy samples. Method: In the present study, murine liver biopsies were performed and stored under various conditions: snap-freezing, RNAlater and Allprotect. Yield was compared to fresh biopsy tissue. Results: Fresh tissue generated the highest yield of RNA while samples subjected to the snap-freezing generated the lowest yield of RNA. Preservation in RNAlater yielded higher quantities of RNA than storage in Allprotect, particularly with larger biopsy samples. There was a non-significant trend toward improved RNA quality with RNAlater (p?=?0.35). DNA and protein yield were similar with RNAlater and Allprotect under a number of handling condition. Errors in tissue handling such as delays in tissue submersion or freezing did not significantly affect tissue yields in either preservation solution. Tissue yield was unchanged with up to three freeze-thaw cycles in both solutions. Biopsy size (5 vs 2?mm) and width (15 vs 18?g) had a marked effect on tissue yield. Conclusion: Ideally 5-mm biopsies with 15-gauge needles should be used to maximize yield. RNAlater provided higher RNA yield with similar yields of DNA and protein and was notably cheaper and easier to handle.