Comparison of fecal DNA extraction kits for the giant panda (Ailuropoda
melanoleuca) by short tandem repeat (STR) genotype analysis
Abstract
Genetic analysis of short tandem repeat (STR) loci using non-invasive
fecal samples is currently the most widely used method in genetic
surveys of giant pandas (Ailuropoda melanoleuca). However, Low-quality
fecal DNA obtained from fecal samples may affects the correctness of
short tandem repeat (STR) genotyping results and poses a challenge to
accurately identify individuals. The aim of this study was thus to
compare the efficiency of DNA extraction kits in obtaining high-quality
fecal host DNA from giant panda fecal samples. In the present study, six
commercial kits widely used in fecal DNA extraction, the QIAamp Fast DNA
Stool Mini Kit (Q kit), Beaver Beads™ Stool DNA Kit (H kit), Mag-MK Soil
& Stool Genome DNA Extraction kit (S kit), Magnetic Soil And Stool DNA
Kit (T kit), E.Z.N.A™ Mag-Bind Stool DNA Kit (O kit) and Mag Beads Fast
DNA Kit for Feces (M kit), were compared to improve the efficiency of
STR genotyping of giant panda fecal DNA samples. Fecal DNA concentration
and purity were measured, and STR genotyping was performed using blood
and fecal DNA from captive giant pandas to compare the genotype matches
at 11 STR loci. Our results show that the most efficient extraction kits
were the Q and T kits, and the Q kit had a greater ability to remove PCR
inhibitors than other kits. However, different STR genotyping systems
require different DNA extraction kits in order to achieve the highest
possible genotyping correctness.