title: Comparison of three methods of DNA extraction from peripheral blood mononuclear cells and lung fragments of equines.
abstract: 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 beta-actin, and cost. Thirty-four 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((R)) 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 A. We conclude that Protocol A should be used for PCR diagnosis from lung fragment samples, while Protocol B should be used for PBMC.
authors: Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brasil. elmaira27@yahoo.com.br
authors: E M
authors: EM
authors: Santos
authors: J F R
authors: JF
authors: Paula
authors: P M C
authors: PM
authors: Motta
authors: M B
authors: MB
authors: Heinemann
authors: R C
authors: RC
authors: Leite
authors: J P A
authors: JP
authors: Haddad
authors: H L
authors: HL
authors: Del Puerto
authors: J K P
authors: JK
authors: Reis
mesh_terms: D000818:Animals; D004247:DNA / Q000302:isolation & purification*; D006736:Horses; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008168:Lung / Q000378:metabolism*; D016133:Polymerase Chain Reaction
chemical_list: D004247:DNA