Back to Search Start Over

Optimizing nucleic acid extraction from thyroid fine-needle aspiration cells in stained slides, formalin-fixed/paraffin-embedded tissues, and long-term stored blood samples

Authors :
Magnus R. Dias-da-Silva
Fernando A. Soares
Janete M. Cerutti
Teresa S. Kasamatsu
Mirian G. Cardoso
Ilda S. Kunii
Susan C. Lindsey
Gisele Giannocco
João Roberto Maciel Martins
Maria Clara C. Melo
Michelle Y. Harada
Marina M. L. Kizys
Rui M. B. Maciel
Marlyn Z. Montoya
Source :
Arquivos Brasileiros de Endocrinologia & Metabologia v.56 n.9 2012, Arquivos Brasileiros de Endocrinologia & Metabologia, Sociedade Brasileira de Endocrinologia e Metabologia (SBEM), instacron:SBEM
Publication Year :
2012
Publisher :
FapUNIFESP (SciELO), 2012.

Abstract

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. Arq Bras Endocrinol Metab. 2012;56(9):618-26

Details

ISSN :
00042730
Volume :
56
Database :
OpenAIRE
Journal :
Arquivos Brasileiros de Endocrinologia & Metabologia
Accession number :
edsair.doi.dedup.....afd6dd61f23ae6f5e1732faad6d953e1
Full Text :
https://doi.org/10.1590/s0004-27302012000900004