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cje ›› 2011, Vol. 30 ›› Issue (01): 1-6.

• Articles •     Next Articles

Characterization of (GT)n microsatellite in Anodonta woodiana genome.

WANG Gui-ling1, SU Xiang1, LI Jia-le1,2**, BAI Zhi-yi1   

  1. 1Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Shanghai Ocean University, Ministry of Education, Shanghai 201306, China|2Aquaculture Division, EInstitute of Shanghai Universities, Shanghai 201306, China
  • Online:2011-01-08 Published:2011-01-08

Abstract: The method of magnetic-bead hybridization enrichment was used to isolate the microsatellite loci of Anodonta woodiana. Genomic DNA was digested with restriction enzyme Sau3A1. The fragments were hybridized with biotin-labeled (GT)15 probe and captured by streptavidin-coated magnetic beads. The target fragments were inserted to PGEM-T vectors, and positive clones were obtained through PCR screening the library. By isolating the (GT) microsatellite in A. woodiana genome, 180 positive clones were sequenced, 37 sequences contained microsatellite having at least 5 times repeats numbers (20.6% of white colonies screened). From the microsatellites sequence, the two bases repeat arrays were most abundant, accounting for 86.4%, among which, 32.0% were perfect repeats, 10.0% imperfect repeats, 8.0% compound repeats, and 50.0% the combination of imperfect repeats and compound repeats. The largest repeat number of perfect type microsatellite was 65. This allowed us to design 30 pairs of primers from 37 microsatellite sequences. As a result, 11 pairs were screened and used successfully to amplify polymorphic fragments.

Key words: Prorocentrum, Harmful algal bloom, Morphology, Taxonomy