Comparison of three methods of DNA extraction from peripheral blood mononuclear cells and lung fragments of equines.
EM Santos, JF Paula, PM Motta… - Genetics and …, 2010 - europepmc.org
EM Santos, JF Paula, PM Motta, MB Heinemann, RC Leite, JP Haddad, HL Del Puerto…
Genetics and molecular research: GMR, 2010•europepmc.orgWe 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 …
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 …
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.
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