川党参的基因组DNA提取与ISSR引物筛选

目的 筛选最适党参基因组DNA的提取方法及ISSR (inter-simple sequence repeat)引物,为研究党参种植资源遗传多样性及DNA鉴定奠定基础.方法 以党参嫩叶作为材料,采用常规SDS法、改良CTAB法和试剂盒法3种提取方法基因组DNA,用优化的ISSR-PCR反应对100条引物进行筛选.结果 以党参嫩叶为材料提取基因组DNA,改良CTAB法效果最佳.从公布的100条ISSR引物中筛选出10条引物,能扩增条带清晰、稳定性好、多态性高的DNA条带.结论 以党参嫩叶为材料提取基因组DNA进行ISSR分子标志研究遗传多样性时应采用改良CTAB法,筛选出合适的引物在ISSR分子标志中很重要.

Genomic DNA extracting and ISSR primer screening of Codonopsis tangshen

Objective In order to study the genetic diversity and DNA identification of Codonopsis pilosula , DNA extracting methods and primers screening were explored. Methods The genomic DNA was extracted from the young leaves using the traditional SDS method, modified CTAB method and DNA isolation kit. The optimized ISSR-PCR reaction was used to screen 100 primers. Results The modified CTAB method was better in extracting genomic DNA from the young leaves. Ten primers were selected to amplify the DNA bands with clear, stable and high polymorphism after screening 100 ISSR primers. Conclusion The modified CTAB method and ten primers selected could be used in the study of genetic diversity by ISSR molecular markers.