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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
- 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
- Subjects :
- Tissue Fixation
Endocrinology, Diabetes and Metabolism
Biopsy, Fine-Needle
Thyroid Gland
Polymerase Chain Reaction
DNA sequencing
chemistry.chemical_compound
blood
medicine
nucleic acid extraction
FFPE tissue
Humans
Thyroid Neoplasms
Electrophoresis, Agar Gel
Paraffin Embedding
Staining and Labeling
medicine.diagnostic_test
biology
RNA
DNA
General Medicine
Proteinase K
Isolation (microbiology)
Molecular biology
Fine-needle aspiration
chemistry
FNA
biology.protein
Nucleic acid
RNA extraction
Subjects
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