Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samples

Nucleic-acid based methods for bacterial identification are extremely useful in diagnostic applications due to their specificity and sensitivity. However, they require an optimal purification of the target molecules. As part of the development of a new diagnostic method for the detection of bacteria...

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Autores principales:	García-Nogales,Paula, Serrano,Alicia, Secchi,Sonia, Gutiérrez,Serafín, Arís,Anna
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2010
Materias:	bacteria milk purification RNA
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500019
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spelling	oai:scielo:S0717-345820100005000192011-05-24Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samplesGarcía-Nogales,PaulaSerrano,AliciaSecchi,SoniaGutiérrez,SerafínArís,Anna bacteria milk purification RNA Nucleic-acid based methods for bacterial identification are extremely useful in diagnostic applications due to their specificity and sensitivity. However, they require an optimal purification of the target molecules. As part of the development of a new diagnostic method for the detection of bacterial RNA in cow milk, we have compared four commercially available RNA extraction kits for the isolation of bacterial RNA from spiked UHT milk samples. The kits were compared in terms of extraction efficiency and RNA purity using two bacterial species, the Gram negative Escherichia coli and the Gram positive Staphylococcus aureus. Two kits are based in silica-matrix extraction, and the other two in the guanidinium thiocyanate-phenol-chloroform extraction. In our hands, the RNeasy Protect Bacteria Mini kit from QIAGEN was the best performing in terms of RNA yield, quality, reproducibility and consumable needs, under the conditions here described.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.13 n.5 20102010-09-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500019en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	bacteria milk purification RNA
spellingShingle	bacteria milk purification RNA García-Nogales,Paula Serrano,Alicia Secchi,Sonia Gutiérrez,Serafín Arís,Anna Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samples
description	Nucleic-acid based methods for bacterial identification are extremely useful in diagnostic applications due to their specificity and sensitivity. However, they require an optimal purification of the target molecules. As part of the development of a new diagnostic method for the detection of bacterial RNA in cow milk, we have compared four commercially available RNA extraction kits for the isolation of bacterial RNA from spiked UHT milk samples. The kits were compared in terms of extraction efficiency and RNA purity using two bacterial species, the Gram negative Escherichia coli and the Gram positive Staphylococcus aureus. Two kits are based in silica-matrix extraction, and the other two in the guanidinium thiocyanate-phenol-chloroform extraction. In our hands, the RNeasy Protect Bacteria Mini kit from QIAGEN was the best performing in terms of RNA yield, quality, reproducibility and consumable needs, under the conditions here described.
author	García-Nogales,Paula Serrano,Alicia Secchi,Sonia Gutiérrez,Serafín Arís,Anna
author_facet	García-Nogales,Paula Serrano,Alicia Secchi,Sonia Gutiérrez,Serafín Arís,Anna
author_sort	García-Nogales,Paula
title	Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samples
title_short	Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samples
title_full	Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samples
title_fullStr	Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samples
title_full_unstemmed	Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samples
title_sort	comparison of commercially-available rna extraction methods for effective bacterial rna isolation from milk spiked samples
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2010
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500019
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Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samples

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