A new method for DNA extraction from feces and hair shafts of the South China tiger (Panthera tigris amoyensis).
It is commonly known that tigers (Panthera tigris) groom themselves by licking their coats, which leads to an abundance of hairs in their feces. These hairs are designated specially as "fecal hairs". In our study, in order to explore fecal hairs potential as a DNA source for genetic analysis, 55 fecal hair samples were collected from 23 captive South China tigers (P. t. amoyensis). According to the amplification of mitochondrial primers loop F and loop R, DNA quality of noninvasive samples were grouped into three grades: grade I-the highest-quality DNA, grade II--high-quality DNA, and grade III--poor-quality DNA. No failed amplifications on microsatellite primers and only 0.27% genotyping errors occurred with grade I fecal hair DNA, as compared with 9.4% failed amplifications on microsatellite primers and 9.5% genotyping errors with grade II fecal hair DNA. It was found that 25.45% of fecal hair DNA was grade I and 65.45 and 10.00% of fecal hair DNA were grades II and III, respectively, as compared with 4.35% grade I fecal DNA and 34.78 and 60.87% grades II and III fecal DNA, respectively. Thus, higher-quality DNA can be extracted from fecal hairs than feces. In addition, DNA could be extracted from hair shafts of tigers and a minimum of 2000 hair shafts were required for visible DNA bands on a 1% agarose gel. These findings demonstrate that fecal hairs may serve as a convenient and reliable genomic DNA source for genotype analysis.
Zhang
Wenping
W
Key Laboratory of Bio-resource and Eco-environment (Ministry of Education), College of Life Science, Sichuan University, Chengdu, Sichuan, People's Republic of China.
Zhang
Zhihe
Z
Xu
Xiao
X
Wei
Kun
K
Wang
Xiaofang
X
Liang
Xu
X
Zhang
Liang
L
Shen
Fujun
F
Hou
Rong
R
Yue
Bisong
B
D000818:Animals; D016172:DNA Fingerprinting / Q000379:methods; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000302:isolation & purification* / Q000592:standards; D005243:Feces; D005838:Genotype; D006120:Grooming; D006197:Hair; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D046010:Tigers / Q000235:genetics*
D017931:DNA Primers; D004272:DNA, Mitochondrial