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Artigo Acesso aberto Produção Nacional Revisado por pares
BIBTEX RIS

Methodology Comparison of three methods of DNA extraction from peripheral blood mononuclear cells and lung fragments of equines

2010; Research Foundation of Ribeirão Preto; Volume: 9; Issue: 3 Linguagem: Inglês

10.4238/vol9-3gmr818

ISSN

1676-5680

Autores

E.M. Santos, J.F.R. Paula, Pedro Moreira Couto Motta, Marcos Bryan Heinemann, R.C. Leite, João Paulo Amaral Haddad, Helen L. Del Puerto, Jenner Karlisson Pimenta dos Reis,

Tópico(s)

Effects of Environmental Stressors on Livestock

Resumo

We compared three different protocols for DNA extraction from horse peripheral blood mononuclear cells (PBMC) and lung fragments, determining average final DNA concentration, purity, percentage of PCR amplification using β-actin, and cost.Thirtyfour samples from PBMC, and 33 samples from lung fragments were submitted to DNA extraction by three different protocols.Protocol A consisted of a phenol-chloroform and isoamylic alcohol extraction, Protocol B used alkaline extraction with NaOH, and Protocol C used the DNAzol ® reagent kit.Protocol A was the best option for DNA extraction from lung fragments, producing high DNA concentrations, with high sensitivity in PCR amplification (100%), followed by Protocols C and B. On the other hand, for PBMC samples, Protocol B gave the highest sensitivity in PCR amplification (100%), followed by Protocols C and ©FUNPEC-RP www.funpecrp.com.

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