Fosmid Library Construction And Optimal Parameter Selection For WGP Physical Map Construction Of Yesso Scallop

The producing areas of yesso scallop (Patinopecten yessoensis) are mainlylocated in northern Japan, eastern Russian and northern Korean Peninsula coastalareas. Its good quality made it become one of the most important marine economicshellfish in China after its introduction to China in1982, and it plays an importantrole in mariculture industry. With the development of next generation sequencing(NGS) technology, characterize the genetic background of species has graduallybecome simple, fast, and low cost. Recent studies of yesso scallop have focused ongenomics deeply. In order to cope with the whole genome sequence assembly ofYessoscallop, a fosmid library covering3.3fold of the genome was constructed forYessoscallop, which was an important part in the whole-genome project. This librarywill not only assist the assembly of the genome, construction of physical map andmaps integration, but also provide recources for map-based gene cloning and itslocalization on chromosomes.In this study, a novel physical mapping technology based on next-generationsequencing, whole genome profiling (WGP), to construct clone contigs with part ofthe clones from the Fosmid library of yesso scallop. The optimum strategy for clonespooling was preliminarily identified, that is, clones in eight384-microwell plates weremixed for a super pool. Using digested tags generated from two kinds of type IIBrestriction endonucleases named BsaXI and FspEI,We could decode mono-clone successfully in the mixed pool and the decodingrate can be up to88%. Meanwhile we carried out preliminary assembly of decodedclones. We demonstrate that WGP maps can be generated for complex bivalvegenomes，and will be further used for the assembling of whole genome sequence.