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|Title: ||Comparison of three DNA extraction methods for Mycobacterium bovis, Mycobacterium tuberculosis and Mycobacterium avium subsp. avium|
|Authors: ||Amaro, Ana|
|Issue Date: ||2008|
|Abstract: ||Aims: To compare three methods for DNA extraction from Mycobacterium
bovis, Mycobacterium tuberculosis and Mycobacterium avium subsp. avium.
Methods and Results: The DNA was extracted from mycobacterial cultures
using enzymatic extraction, combined bead beating and enzymatic extraction
and cetyltrimethylammonium bromide (CTAB) extraction. The yield and quality
of DNA were compared by spectrophotometry, agarose gel electrophoresis,
restriction endonuclease analysis and PCR. The combined bead beating and
enzymatic extraction method yielded more DNA. However, that method produced
some sheared DNA, visible either by agarose gel electrophoresis or by
restriction endonuclease analysis. All methods were appropriate for PCR amplification
of a 123 bp fragment of IS6110 in M. bovis and M. tuberculosis, and of
a 1700 bp fragment of FR300 region in M. avium avium.
Conclusions: Combined bead beating and enzymatic extraction method was
the most efficient and easy method for extracting DNA from bacteria of the
M. tuberculosis complex.
Significance and Impact of the Study: The results reveal important differences
among the DNA extraction methods for mycobacteria, which are relevant for
the success of further downstream molecular analysis.|
|Appears in Collections:||MVT - Publicações - Artigos em Revistas Internacionais Com Arbitragem Científica|
ICAAM - Publicações - Artigos em Revistas Internacionais Com Arbitragem Científica
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