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A Modified Enrichment Method to Construct Microsatellite Libraryfrom Plateau Pika Genome (Ochotona curzoniae)
Geng Jianing; Li Kexin; Zhang Yanming; Hu Songnian
2010
Source PublicationGenomics, Proteomics & Bioinformatics
Volume8Issue:1Pages:72-76
AbstractA microsatellite-enriched library of plateau pika (Ochotona curzoniae) was constructed according to the strongaffinity between biotin and streptavidin. Firstly, genomic DNA was fragmented by ultrasonication, which is a majorimprovement over traditional methods. Linker-ligated DNA fragments were hybridized with biotinylated microsatelliteprobes, and then were subjected to streptavidin-coated magnetic beads. PCR amplificationwasperformedto obtain double-stranded DNA fragments containing microsatellites. Ligation and transformation werecarried out by using the pGEM-T Vector System I and Escherichia coli DH10B competent cells. Sequencing resultsshowed that 80.2% of clones contained microsatellite repeatmotif. Several modifications make this protocoltime-efficient and technically easier than the traditional ones; particularly,composition and relative abundance ofmicrosatellite repeats in plateau pika genome were truly represented through the optimized PCR conditions. Thismethod has also been successfully applied to construct microsatellite-enriched genomic libraries of Chinese hamster(Cricetulus griseus) and small abalone [Haliotis diversicolor (Reeve)] with high rates of positive clones,demonstrating its feasibility and stability.
KeywordMicrosatellite Enriched Library Magnetic Bead Ultrasonication
Indexed ByCSCD
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Document Type期刊论文
Identifierhttp://210.75.249.4/handle/363003/1492
Collection中国科学院西北高原生物研究所
Recommended Citation
GB/T 7714
Geng Jianing,Li Kexin,Zhang Yanming,et al. A Modified Enrichment Method to Construct Microsatellite Libraryfrom Plateau Pika Genome (Ochotona curzoniae)[J]. Genomics, Proteomics & Bioinformatics,2010,8(1):72-76.
APA Geng Jianing,Li Kexin,Zhang Yanming,&Hu Songnian.(2010).A Modified Enrichment Method to Construct Microsatellite Libraryfrom Plateau Pika Genome (Ochotona curzoniae).Genomics, Proteomics & Bioinformatics,8(1),72-76.
MLA Geng Jianing,et al."A Modified Enrichment Method to Construct Microsatellite Libraryfrom Plateau Pika Genome (Ochotona curzoniae)".Genomics, Proteomics & Bioinformatics 8.1(2010):72-76.
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