A sensitive and specific multiplex PCR approach for sex identification of ursine and tremarctine bears suitable for non-invasive samples.
We report a new approach for molecular sex identification of extant Ursinae and Tremarctinae bears. Two Y-specific fragments (SMCY and 318.2) and one X-specific fragment (ZFX) are amplified in a multiplex PCR, yielding a double test for male-specific amplification and an internal positive control. The primers were designed and tested to be bear-specific, thereby minimizing the risk of cross-amplification in other species including humans. The high sensitivity and small amplicon sizes (100, 124, 160 base pairs) facilitate analysis of non-invasively obtained DNA material. DNA from tissue and blood as well as from 30 non-invasively collected hair and faeces yielded clear and easily interpretable results. The fragments were detected both by standard gel electrophoresis and automated capillary electrophoresis.
LOEWE Biodiversity and Climate Research Centre (BiK-F), Senckenberg Gesellschaft für Naturforschung, Senckenberganlage 25, 60325, Frankfurt am Main, Germany. tobias.bidon@senckenberg.de
Tobias
T
Bidon
Christiane
C
Frosch
Hans G
HG
Eiken
Verena E
VE
Kutschera
Snorre B
SB
Hagen
Siv G
SG
Aarnes
Steven R
SR
Fain
Axel
A
Janke
Frank
F
Hailer
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D019075:Electrophoresis, Capillary; D005243:Feces / Q000737:chemistry; D005260:Female; D005819:Genetic Markers / Q000235:genetics*; D006197:Hair / Q000737:chemistry; D051741:Kruppel-Like Transcription Factors / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012732:Sex Determination Analysis / Q000379:methods*; D013045:Species Specificity; D001503:Ursidae / Q000235:genetics*; D014998:Y Chromosome / Q000235:genetics*
D017931:DNA Primers; D005819:Genetic Markers; D051741:Kruppel-Like Transcription Factors; C062076:zinc finger protein, X-linked