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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 :
Kizys MM
Cardoso MG
Lindsey SC
Harada MY
Soares FA
Melo MC
Montoya MZ
Kasamatsu TS
Kunii IS
Giannocco G
Martins JR
Cerutti JM
Maciel RM
Dias-da-Silva MR
Source :
Arquivos brasileiros de endocrinologia e metabologia [Arq Bras Endocrinol Metabol] 2012 Dec; Vol. 56 (9), pp. 618-26.
Publication Year :
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.<br />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.<br />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.<br />Conclusion: The techniques herein applied to isolate nucleic acids allowed further reliable molecular analyses.

Details

Language :
English
ISSN :
1677-9487
Volume :
56
Issue :
9
Database :
MEDLINE
Journal :
Arquivos brasileiros de endocrinologia e metabologia
Publication Type :
Academic Journal
Accession number :
23329184
Full Text :
https://doi.org/10.1590/s0004-27302012000900004