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

C.G. Oliveira¹, R.A. Martinez^1,2 and F.A. Gaiotto^1,3
1Programa de Pós-Graduação em Genética e Biologia Molecular, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brasil
²Departamento de Filosofia e Ciências Humanas, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brasil
³Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brasil
Corresponding author: R.A. Martinez
E-mail: cebus@yahoo.com

Genet. Mol. Res. 6 (3): 657-666 (2007)
Received June 24, 2007
Accepted August 10, 2007
Published September 30, 2007

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 conservation. Chaetomys subspinosus (Rodentia: Erethizontidae) is a secretive arboreal porcupine endemic to certain ecosystems of the Brazilian Atlantic 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 adapting a DNA extraction protocol traditionally used for plants. Digestion using proteinase K was followed by protein precipitation with CTAB, a chloroform-isoamyl alcohol cleaning and DNA precipitation with isopropyl alcohol. This protocol supplies good-quality DNA for genetic analysis with molecular markers based on PCR.

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