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Extraction, PCR amplification, and sequencing of mitochondrial DNA from scent mark and feces in the giant panda
Author(s) -
Ding Bo,
Zhang Yaping,
Ryder Oliver A.
Publication year - 1998
Publication title -
zoo biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.5
H-Index - 54
eISSN - 1098-2361
pISSN - 0733-3188
DOI - 10.1002/(sici)1098-2361(1998)17:6<499::aid-zoo3>3.0.co;2-p
Subject(s) - biology , mitochondrial dna , polymerase chain reaction , cytochrome b , feces , dna , microbiology and biotechnology , gene , oligonucleotide , dna sequencing , genetics , dna extraction
To expand the feasibility of applying simple, efficient, non‐invasive DNA preparation methods using samples that can be obtained from giant pandas living in the wild, we investigated the use of scent markings and fecal samples. Giant panda–specific oligonucleotide primers were used to amplify a portion of the mitochondrial DNA control region as well as a portion of the mitochondrial DNA cytochrome b gene and tRNA Thr gene region. A 196 base pair (bp) fragment in the control region and a 449 bp fragment in the cytochrome b gene and tRNA Thr gene were successfully amplified. Sequencing of polymerase chain reaction (PCR) products demonstrated that the two fragments are giant panda sequences. Furthermore, under simulated field conditions we found that DNA can be extracted from fecal samples aged as long as 3 months. Our results suggest that the scent mark and fecal samples are simple, efficient, and easily prepared DNA sources. Zoo Biol 17:499–504, 1998. © 1998 Wiley‐Liss, Inc.