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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 13, No. 5, 2010
Bioline Code: ej10063
Full paper language: English
Document type: Note
Document available free of charge

Electronic Journal of Biotechnology, Vol. 13, No. 5, 2010

 en Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samples
García-Nogales, Paula; Serrano, Alicia; Secchi, Sonia; Gutiérrez, Serafín & Arís, Anna

Abstract

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 check for this species in other resources and the Gram positive Staphylococcus aureus check for this species in other resources . 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.

Keywords
bacteria, milk, purification, RNA.

 
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