Título
Simple and inexpensive DNA extraction protocol for studying the bacterial composition of sludges used in microbial fuel cells
Autor
BLONDY BEATRIZ CANTO CANCHE
Nivel de Acceso
Acceso Abierto
Resumen o descripción
Bacteria oxidize organic matter and nutrients to produce
electric energy in microbial fuel cells (MFC) - a technology of increasing
importance because of its sustainability. To improve the performance
of MFCs, it is necessary not only to gain a better understanding of
MFC engineering designs, but also to improve the understanding of
the composition of the microbial communities in MFCs. Fast and
efficient DNA extraction protocols that are suitable for extracting
diverse bacterial genomes are necessary to identify the bacterial
diversity present in MFCs and to further monitor the dynamic changes
of microbial communities. This study focused on testing different direct
cell lysis protocols to extract DNA from a microbial sludge harvested from an MFC. The protocol that achieved the best results was based on
a previous study, but was modified by eliminating a chaotropic salt and
the special columns used for nucleic acid purification. The efficiency of
this less expensive and more straightforward protocol was confirmed
by PCR amplification of the 16S rRNA gene and denaturing gradient
gel electrophoresis analysis, which confirmed the extraction of multiple
genomes. The sequences of 10 clones revealed the presence of phyla,
Proteobacteria, Firmicutes and Actinobacteria, comprising both Gramnegative
and Gram-positive bacteria. Some of these bacteria were
identified at the genus level, e.g., Clostridium, Pseudoxanthomonas,
Tistrella, and Enterobacter; these genera have been described in active
sludges from wastewater treatment, supporting the congruency of
our results. Therefore, this protocol is a useful tool for analysis of the
bacteria responsible for energy production in MFCs.
Fecha de publicación
4 de febrero de 2013
Tipo de publicación
Artículo
Recurso de información
Formato
application/pdf
Repositorio Orígen
Repositorio Institucional CICY
Descargas
1532