Paper Info
Title
A modified enrichment method to construct microsatellite library from plateau pika genome (Ochotona curzoniae).
Abstract
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
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 Geng160.89
Kexin Li200.68
kexin300.34
Songnian Hu48716.83
yanming500.68
yanming600.68
songnian751.87
hu841.48