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1 Bulletin of the Veterinary Institute in Puławy

1 2009

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Optimisation of the DNA-extraction method from foal faeces for the diagnosis of Rhodococcus equi infections using PCR

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Zietek-Barszcz A. , Grzadzki Z.

Bulletin of the Veterinary Institute in Puławy

2009

tom 53

nr 3

The aim of the study was to compare the effectiveness of selected DNA extraction methods for the detection of Rhodococcus equi in foal faeces using PCR. Four different nucleic acid extraction methods were compared, which were based mainly on the use of proteolitic enzymes. Moreover, the commercial kit for DNA isolation from faeces was used. In two of the methods additional components, including CTAB detergent and powdered glass, were introduced. The PCR results showed that traditional DNA extraction methods involving proteolitic enzymes and the commercial kit were ineffective in terms of preparing the DNA matrix amplified by PCR. The only method useful for preparing the pure DNA matrix free from inhibitors of the enzymatic reaction was that with the use of powdered glass.