Knowledge Management System of Northwest Institute of Plateau Biology, CAS
A Modified Enrichment Method to Construct Microsatellite Libraryfrom Plateau Pika Genome (Ochotona curzoniae) | |
Geng Jianing ; Li Kexin ; Zhang Yanming ; Hu Songnian | |
2010 | |
发表期刊 | Genomics, Proteomics & Bioinformatics
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摘要 | A 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. |
文献类型 | 期刊论文 |
条目标识符 | http://210.75.249.4/handle/363003/35707 |
专题 | 中国科学院西北高原生物研究所 |
推荐引用方式 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, Geng Jianing.A Modified Enrichment Method to Construct Microsatellite Libraryfrom Plateau Pika Genome (Ochotona curzoniae).Genomics, Proteomics & Bioinformatics,2010,8(1):72-76,2010. |
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. |
MLA | Geng Jianing,et al."A Modified Enrichment Method to Construct Microsatellite Libraryfrom Plateau Pika Genome (Ochotona curzoniae)".Genomics, Proteomics & Bioinformatics (2010). |
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