Comparison of Two Methods for the Isolation of Genomic DNA from Cyathostomin Adult Parasites

Yıl 2017, Cilt: 43 Sayı: 2, 146 - 148, 09.06.2017

Juliana Bana Ishii Amilcar Arenal Yanise Chicoy Marcelo Beltrão Molento

https://doi.org/10.16988/iuvfd.320203

Öz

Cyathostomins are the
most common and important group of large intestine nematodes, infecting horses
worldwide. The current control strategy is associated with the development of
anthelmintic resistance, which has been reported worldwide. Therefore,
experiments with this family of parasites have become progressively important
to provide their monitoring and control strategies. The aim of the present
study was to propose a faster and more economic assay for isolation of genomic
DNA from the adult stage of Cyathostomin parasites than reported. Adult
parasites were collected from a single horse from a farm in São José dos
Pinhais, PR, Brazil, and were identified. Genomic DNA was isolated from ten
individual female adult parasites using a standardized procedure developed.
Then, extraction from ten individual female was carried out by another DNA
extraction method. DNA concentration from both methods were measured and
compared. We obtained a good DNA quality with this standardized procedure. As a
result of this analysis, we propose a modified phenol-chloroform method, which
will contribute to assays that require DNA extraction from adult worms for
genomic DNA sequences of cyathostomin, or species-specific identification.

Anahtar Kelimeler

DNA extraction, genomic material, horse, Strongylida

Comparison of Two Methods for the Isolation of Genomic DNA from Cyathostomin Adult Parasites

Öz

Cyathostomins are the most common and important group of large intestine nematodes, infecting horses worldwide. The current control strategy is associated with the development of anthelmintic resistance, which has been reported worldwide. Therefore, experiments with this family of parasites have become progressively important to provide their monitoring and control strategies. The aim of the present study was to propose a faster and more economic assay for isolation of genomic DNA from the adult stage of Cyathostomin parasites than reported. Adult parasites were collected from a single horse from a farm in São José dos Pinhais, PR, Brazil, and were identified. Genomic DNA was isolated from ten individual female adult parasites using a standardized procedure developed. Then, extraction from ten individual female was carried out by another DNA extraction method. DNA concentration from both methods were measured and compared. We obtained a good DNA quality with this standardized procedure. As a result of this analysis, we propose a modified phenol-chloroform method, which will contribute to assays that require DNA extraction from adult worms for genomic DNA sequences of cyathostomin, or species-specific identification.

Anahtar Kelimeler

DNA extraction, genomic material, horse, Strongylida

Kaynakça

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