Isolation And Development Of Microsatellites From Ramie (Boehmeria Nivea L. Gaud)

The development of Ramie breeding by molecular markers is going rapid, In the study, the DNA database search method and the magnetic beads probes hybridization enrichment separation by 41 ramie microsatellite sequence, Designed 41 Pairs of microsatellites sequence by Primer premier5.0and SSRHunterl.3. The main results were:1) From the ramie (Boehmeria nivea L. Gaud) EST database in NCBI,19 sequences including SSR sequence from 320 EST sequences were screened. SSR primers developed from this database.19 pairs of SSR primers were designed based on this sequences.2) AFLP and magnetic beads enrichment were combined to detect SSR primers. The fragments of AFLP pre-amplifacation were bond with linkers, Microsatellite sequence library construction, the expansion of positive screening method cloning, those sequences were hybridized with biotin-labeled simple sequence repeat probes (AG) 15. Microsatellites were efficiently isolated by using magnetic beads coated with streptavidin. Genomic library contained the selected microsatellite DNA was construeted.28 positive clones were isolated with PCR method.6 sequences were deleted because of the short sequence near the SSR site and 22 pairs of SSR primers were successfully designed.