Comparison of three guanidinine thiocyanate RNA extraction methods applied to shellfish extract for enteric virus detection
Paper Details
Comparison of three guanidinine thiocyanate RNA extraction methods applied to shellfish extract for enteric virus detection
Abstract
Human enteric viruses are transmitted through water and food. It’s therefore necessary to include the research of these agents in the food and water sanitary quality analysis. Most of these viruses are not culturable. Gene amplification methods such as NASBA and RT-PCR are used for routine analysis. These methods of detection are possible only if there is an adapted procedure for extracting nucleic acids especially RNA. We proposed here to evaluate the quality of the three RNA extraction protocols using the guanidine thiocyanate. Two of the considered extraction methods are based on the work of Chomczynski and Sacchi (method I and method II). A third one which is a modification in order to reduce the working time was also applied (method III). Extraction tests were performed on mussel extract after concentration with PEG / NaCl . The quality of extraction is evaluated by spectrophotometry on the criteria “yield ” and ” purity” . The results show that method I and method II are equivalent and suitable and the method III was not appropriate for this type of samples.
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