Arquivos brasileiros de endocrinologia e metabologia · 2012
Optimizing nucleic acid extraction from thyroid fine-needle aspiration cells in stained slides, formalin-fixed/paraffin-embedded tissues, and long-term stored blood samples.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
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Kizys Marina M L, Cardoso Mirian G, Lindsey Susan C, Harada Michelle Y, Soares Fernando A, Melo Maria Clara C, Montoya Marlyn Z, Kasamatsu Teresa S, Kunii Ilda S, Giannocco Gisele, Martins João Roberto M, Cerutti Janete M, Maciel Rui M B, Dias-da-Silva Magnus R
Аннотация
OBJECTIVE: Adequate isolation of nucleic acids from peripheral blood, fine-needle aspiration cells in stained slides, and fresh and formalin-fixed/paraffin-embedded tissues is crucial to ensure the success of molecular endocrinology techniques, especially when samples are stored for long periods, or when no other samples can be collected from patients who are lost to follow-up. Here, we evaluate several procedures to improve current methodologies for DNA (salting-out) and RNA isolation.
MATERIALS AND METHODS: We used proteinase K treatment, heat shock, and other adaptations to increase the amount and quality of the material retrieved from the samples.
RESULTS: We successfully isolated DNA and RNA from the samples described above, and this material was suitable for PCR, methylation profiling, real-time PCR and DNA sequencing.
CONCLUSION: The techniques herein applied to isolate nucleic acids allowed further reliable molecular analyses.
MATERIALS AND METHODS: We used proteinase K treatment, heat shock, and other adaptations to increase the amount and quality of the material retrieved from the samples.
RESULTS: We successfully isolated DNA and RNA from the samples described above, and this material was suitable for PCR, methylation profiling, real-time PCR and DNA sequencing.
CONCLUSION: The techniques herein applied to isolate nucleic acids allowed further reliable molecular analyses.