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Comparison of fecal DNA extraction kits for the giant panda (Ailuropoda melanoleuca) by short tandem repeat (STR) genotype analysis
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  • Jie Gao,
  • Chunhai Li,
  • Yitao Wu,
  • Xingyong Zhu,
  • Siqin Liu,
  • Yang Zhang,
  • Huizhong Pang,
  • Jiaheng Li,
  • Jiawen Liu,
  • Wangsheng Zhao,
  • Ye Wang,
  • Jie Kou
Jie Gao
Chengdu Research Base of Giant Panda Breeding
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Chunhai Li
Chengdu Research Base of Giant Panda Breeding
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Yitao Wu
Chengdu Research Base of Giant Panda Breeding
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Xingyong Zhu
Chengdu Research Base of Giant Panda Breeding
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Siqin Liu
Chengdu Research Base of Giant Panda Breeding
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Yang Zhang
Chengdu Research Base of Giant Panda Breeding
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Huizhong Pang
Chengdu Research Base of Giant Panda Breeding
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Jiaheng Li
Chengdu Research Base of Giant Panda Breeding
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Jiawen Liu
Chengdu Research Base of Giant Panda Breeding
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Wangsheng Zhao
Southwest University of Science and Technology School of Life Sciences and Engineering
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Ye Wang
Chengdu Research Base of Giant Panda Breeding
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Jie Kou
Chengdu Research Base of Giant Panda Breeding

Corresponding Author:koujie@panda.org.cn

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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.
30 Nov 2024Submitted to Ecology and Evolution
06 Dec 2024Submission Checks Completed
06 Dec 2024Assigned to Editor
12 Dec 2024Reviewer(s) Assigned
06 Jan 2025Review(s) Completed, Editorial Evaluation Pending
13 Jan 2025Editorial Decision: Revise Minor