Title | ||
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A modified enrichment method to construct microsatellite library from plateau pika genome (Ochotona curzoniae). |
Abstract | ||
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A microsatellite-enriched library of plateau pika (Ochotona curzoniae) was constructed according to the strong affinity between biotin and streptavidin. Firstly, genomic DNA was fragmented by ultrasonication, which is a major improvement over traditional methods. Linker-ligated DNA fragments were hybridized with biotinylated microsatellite probes, and then were subjected to streptavidin-coated magnetic beads. PCR amplification was performed to obtain double-stranded DNA fragments containing microsatellites. Ligation and transformation were carried out by using the pGEM-T Vector System I and Escherichia coli DH10B competent cells. Sequencing results showed that 80.2% of clones contained microsatellite repeat motif. Several modifications make this protocol time-efficient and technically easier than the traditional ones; particularly, composition and relative abundance of microsatellite repeats in plateau pika genome were truly represented through the optimized PCR conditions. This method 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. |
Year | DOI | Venue |
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2010 | 10.1016/S1672-0229(10)60007-1 | Genomics, Proteomics & Bioinformatics |
Keywords | Field | DocType |
microsatellite,enriched library,magnetic bead,ultrasonication | Genome,Polymerase chain reaction,Ochotona curzoniae,Biology,Microsatellite,Genomic library,Genetics,Pika,Molecular biology,Microsatellite Repeat,genomic DNA | Journal |
Volume | Issue | ISSN |
8 | 1 | 1672-0229 |
Citations | PageRank | References |
0 | 0.34 | 0 |
Authors | ||
8 |
Name | Order | Citations | PageRank |
---|---|---|---|
Jianing Geng | 1 | 6 | 0.89 |
Kexin Li | 2 | 0 | 0.68 |
kexin | 3 | 0 | 0.34 |
Songnian Hu | 4 | 87 | 16.83 |
yanming | 5 | 0 | 0.68 |
yanming | 6 | 0 | 0.68 |
songnian | 7 | 5 | 1.87 |
hu | 8 | 4 | 1.48 |