DNA extraction from bristles and quills of Chaetomys subspinosus (Rodentia: Erethizontidae) using a novel protocol

C.G. Oliveira, R.A. Martinez, F.A. Gaiotto
Published September 30, 2007
Genet. Mol. Res. 6 (3): 657-666 (2007)

About the authors
C.G. Oliveira, R.A. Martinez, F.A. Gaiotto

Corresponding author
R.A. Martinez
E-mail: cebus@yahoo.com

ABSTRACT

DNA extraction protocols are as varied as DNA sources. When it comes to endangered species, it is especially important to pay attention to all details that ensure the completion of the study goals and effectiveness in attaining useful data for conserva- tion. Chaetomys subspinosus (Rodentia: Erethizontidae) is a secretive arboreal porcupine endemic to certain ecosystems of the Brazilian At- lantic Forest. A multidisciplinary study (including genetic data) was performed to create a management plan for the conservation of this species. Individuals from natural populations of the States of Bahia,Espírito Santo and Sergipe were sampled. To obtain a reliable and abundant amount of starting material, non-destructive methods were tested, extracting DNA from the bristles and quills that comprise most of this animal’s hide. This method has also been innovative in adapt- ing a DNA extraction protocol traditionally used for plants. Digestion using proteinase K was followed by protein precipitation with CTAВ, a chloroform-isoamyl alcohol cleaning and DNA precipitation with isopropyl alcohol. This protocol supplies good-quality DNA for ge- netic analysis with molecular markers based on PCR.

Key words: Bristle/hair DNA extraction, Chaetomys subspinosus, Threatened species, Molecular markers, Atlantic Forest, Brazil

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