A cost-effective protocol for total DNA isolation from animal tissue

Abstract

Laboratories that perform PCR on a routine basis need to count on reliable DNA isolation methods. In locations where supply of DNA extraction kits depends on importation, having an in-house protocol is desirable. This is also important for laboratories limited by budget constraints. We present a low-cost DNA isolation protocol that incorporates well-known techniques, but that we have adapted to various animal tissues. We tested this protocol on animal blood and muscle, and on cell suspension from skin swabs. The results were comparable, in terms of amount and quality of DNA, to those obtained with two other commercially available methods. DNA retrieved with this protocol has been successfully employed for Sanger sequencing of gene PCR products from animal tissues and blood, as well as for PCR-based diagnosis of chytrid fungus in amphibians and blood parasites in birds.

Two new glassfrogs (Centrolenidae:Hyalinobatrachium) from Ecuador, with comments on the endangered biodiversity of the Andes

“…We sequenced mitochondrial 16S in 37 individuals, including the two new taxa described below, as well as the morphologically similar H. valerioi ( Dunn, 1931 ) and H. aureoguttatum ( Barrera-Rodriguez & Ruiz-Carranza, 1989 ) ( Material S2 ). Extraction, amplification, and sequencing protocols are described in Guayasamin et al (2008) and Peñafiel et al (2019) . The obtained data were compared with homologous sequences from all available species in the genus Hyalinobatrachium and its sister taxon Celsiella ( Guayasamin et al, 2008 ), downloaded from GenBank ( ) and generated mostly by Guayasamin et al (2008) , Castroviejo-Fisher et al (2014) , and Twomey, Delia & Castroviejo-Fisher (2014) .…”

A new species of the genus Noblella (Amphibia: Strabomantidae) from Ecuador, with new information for Noblella worleyae

“…nov. (ZSFQ 050-051). DNA was extracted from muscle or liver tissue following the protocol by Peñafiel et al (2019). Standard polymerase chain reaction (PCR) was performed to amplify a fragment of the mitochondrial gene 16S rRNA, using a combination of the following primers: 16L10, 16H36E, 16L34, 16H47 (Heinicke et al, 2007).…”

Una fantástica nueva especie del grupoPristimantis orceside los Andes sur de Ecuador

“…Para la extracción de ADN se utilizó el protocolo diseñado por Peñafiel et al [18], mediante el cual se extrajo ADN genómico de tejido hepático conservado en etanol. Amplificamos el gen mitocondrial 12S, utilizando una combinación de cebadores (12L29E-F, t-Phe-frog-R, 16H50-R) y protocolos de amplificación descritos en Heinicke et al [19].…”