Development of a Rapid and Efficient Method for Non-Lethal DNA Sampling and Genotyping in Scallops

PLoS One

Xiaoteng Fu¹  Jia Lv¹  Zhenmin Bao¹  Yan Miao¹  Ru Zhang¹  Lingling Zhang¹  Junxia Mao¹  Xiaoli Hu¹  Shi Wang¹  Liping Hu¹  Changsen Sun² 

[1] Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao, China;School of Life Science, Taizhou University, Taizhou, China

关键词: Genotyping;    DNA extraction;    DNA electrophoresis;    Polymerase chain reaction;    Molecular genetics;    Agarose gel electrophoresis;    Gills;    Experimental design;   

DOI  :  10.1371/journal.pone.0068096

学科分类：医学（综合）

来源: Public Library of Science

【 摘 要 】

Non-lethal DNA sampling has long appealed to researchers studying population and conservation genetics, as it does not necessitate removing individuals permanently from their natural environment or destroying valuable samples. However, such an approach has not yet been well established in bivalves. In this study, we demonstrate that the gill represents a good source of tissue for non-lethal sampling in scallops. Removal of a few gill filaments caused no noticeable behavioral abnormalities or increased mortality rates in Zhikong scallop (Chlamys farreri) during a three-month period of observation. To facilitate rapid gill-based DNA extraction, six methods (MA-MF) were designed and evaluated, each requiring less than one hour of processing time. The optimal method was identified as MF, in terms of maintaining DNA integrity and genotyping accuracy. Further optimization of MF method by orthogonal experimental design suggested that the utilization of gills could be limited to 2 mg of sample, which is sufficient for performing up to 20,000 PCR reactions. We also demonstrate the excellent cross-species utility of MF in two additional scallop species, Yesso scallop (Patinopecten yessoensis) and bay scallop (Argopecten irradians). Taken together, our study provides a rapid and efficient approach for applying non-lethal DNA sampling in bivalve species, which would serve as a valuable tool for maintaining bivalve populations and conservation genetics, as well as in breeding studies.

【 授权许可】

CC BY