title: Development of a rapid and efficient method for non-lethal DNA sampling and genotyping in scallops.
abstract: 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.
authors.affiliation: Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao, China.
authors.forename: Junxia
authors.initials: J
authors.lastname: Mao
authors.forename: Jia
authors.initials: J
authors.lastname: Lv
authors.forename: Yan
authors.initials: Y
authors.lastname: Miao
authors.forename: Changsen
authors.initials: C
authors.lastname: Sun
authors.forename: Liping
authors.initials: L
authors.lastname: Hu
authors.forename: Ru
authors.initials: R
authors.lastname: Zhang
authors.forename: Xiaoteng
authors.initials: X
authors.lastname: Fu
authors.forename: Lingling
authors.initials: L
authors.lastname: Zhang
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Hu
authors.forename: Shi
authors.initials: S
authors.lastname: Wang
authors.forename: Zhenmin
authors.initials: Z
authors.lastname: Bao
mesh_terms: D000818:Animals; D004247:DNA / Q000302:isolation & purification*; D005838:Genotype; D005880:Gills / Q000737:chemistry; D058889:Molecular Typing; D049897:Pectinidae / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D013045:Species Specificity
chemical_list: D004247:DNA