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Loss of mRNA template for molecular markers in tissue compared to the method and storage over time
Author(s) -
Jewell Scott,
Tang Yufang,
Jones Susie,
Mohamed Nehad,
Long Susan,
Yearsley Kurtis,
Kaletunc Gonul
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.72.4
Subject(s) - messenger rna , biomedical engineering , computational biology , chemistry , biology , medicine , biochemistry , gene
The degradation in the quality of biospecimens, especially for human tissues, is estimated to have a significant affect on the molecular analysis and outcome of research data. At present it is not well understood whether methods used in the collection, processing, and storage of biospecimens contribute to a lack of quality of the specimens nor whether standard measures of the quality of biospecimens can help to identify biologically similar or dissimilar biospecimens prior to research use. To determine if processes in tissue packaging and length of storage could affect the quality of tissues we developed a study to measure the degradation (loss) of the mRNA for relevant markers in colon mucosa. We packaged snap frozen tissues in routinely used cryovials, vacuum packaging using a gas impermeable wrapping and packaging with argon. The original quantitative level if the tissue mRNA‐template for three mucosal markers was compared to the mRNA level of the same markers in the tissue stored over several months. The quantitative mRNA values were measured using qRT‐PCR. We also measured protein content by image analysis assisted quantitative immunohistochemistry. The results obtained showed an approximate 5% loss of the original template for most markers within one year of storage at ‐80C and no significant difference in packaging methods to this point in time. The implications of the loss of template suggest that while qualitative results from the comparison of tissues stored over varying periods may not be problematic between wide degrees of storage periods, quantitative measures could provide varying results and unqualified interpretation of results if the processing and storage parameters are not understood or improved.