Microsatelliteenriched library construction and polymorphic microsatellite marker isolation in Charybdis japonica
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    Abstract:

    The method of magnetic bead hybridization enrichment was used to screen the microsatellite molecular markers of Charybdis japonica. Genomic DNA of Charybdis japonica was extracted and digested with restriction enzyme Sau3 A I.The fragments of 400-1 200 bp were recycled and purified.DNA fragments which containing microsatellite sequence were screened with(AC)15 oligonucleotide probe and connected to pMD18-T vector.Genomic library contained the microsatellite sequence was constructed.Positive clones were isolated with PCR method and sequencing.After sequence analysis on 369 positive clones randomly picked from 970 colonies,321(86.99%)of the colonies were found to contain microsatellite sequences.Among the 321 microsatellites,80.54% were perfect,15.95% were imperfect and 4.28% turned out to be compound.From the primers designed for the 102 microsatellite loci,65 could amplify expected PCR products and 27 were found to be polymorphic.The results of this work may contribute to future studies of Charybdis japonica molecular genetic breeding.

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LI Jian, LIU Ping, CHEN Ping, GAO Bao-quan. Microsatelliteenriched library construction and polymorphic microsatellite marker isolation in Charybdis japonica[J]. Journal of Fisheries of China,2011,35(1):35~42

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History
  • Received:October 11,2010
  • Revised:November 11,2010
  • Adopted:November 30,2010
  • Online: January 14,2011
  • Published: January 15,2011
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