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Am J Clin Pathol ; 111(6): 748-52, 1999 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10361509

RESUMO

Molecular diagnosis of malignant lymphoma depends on the ability to extract high molecular weight genomic DNA. However, collection, storage, and transportation of frozen tissue is time consuming and expensive. We used a simple, low-cost lysis, storage, and transportation buffer (LST) to maintain clinical tissue samples at room temperature for up to 4 weeks before molecular analysis. Immersion of lymphoid tissue in LST at room temperature for 2 to 4 weeks was compared with snap-freezing in liquid nitrogen followed by storage at -75 degrees C. Southern blot analysis using an immunoglobulin heavy chain JH probe yielded identical results in 5 clonal and 6 nonclonal samples. The DNA recovered from the LST of a 12th sample was too degraded to be analyzed; however, the tissue had large zones of geographic necrosis. We also demonstrated that DNA extracted from tissue stored in LST is suitable for amplification by the polymerase chain reaction. Results from 4 of the snap-frozen and LST samples analyzed for rearrangements at the immunoglobulin heavy chain VDJ locus were identical. LST can be used in a clinical laboratory for storing tissue samples at room temperature up to 4 weeks before molecular analysis.


Assuntos
Citogenética/métodos , DNA/isolamento & purificação , Tecido Linfoide/citologia , Polietilenoglicóis , Polissorbatos , Preservação de Tecido/métodos , Trometamina , Southern Blotting , Eletroforese , Rearranjo Gênico , Genes de Imunoglobulinas , Humanos , Peso Molecular , Reação em Cadeia da Polimerase
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