SSR primers development in non-heading Chinese cabbage by ISSR-PCR and streptavidin-coated magnetic beads adsorption

SSR primers development in non-heading Chinese cabbage by ISSR-PCR and streptavidin-coated magnetic beads adsorption

2006

A:PS

Two methods of ISSR-PCR and streptavidin-coated magnetic beads adsorption were used to isolate microsatellites from genomic DNA of non-heading Chinese cabbage (Brassica campestris subsp. chinensis [Brassica pekinensis]) and a total of 41 SSR primers were successfully developed. SSR primers (IP2 and IP3) were designed on the basis of the determined terminal sequence of nested PCR products in the ISSR-PCR method, with an efficiency of 12% for SSR primer development. As to the method of streptavidin-coated magnetic beads adsorption, firstly, the biotin-labelled SSR probe was hybridized with digested genomic DNA and SSR fragments were enriched by absorption with streptavidin-coated magnetic beads and then they were amplified, cloned and sequenced. On the basis of sequence flanking, microsatellite SSR primers (PL and PR) were designed with an efficiency of 9.6%.

v.33(1):155-157, ACTA HORTICULTURAE SINICA

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