Isolation of high-quality genomic DNA of transgenic Dendrobium plantlets for rapid screen or genetic analysis by PCR amplification and Southern blot is difficult, for the transgenic plantlets grow very slowly and contain high-level polysaccharides. In order to overcome such problems as low yield, degradation, poor PCR amplification and poor restriction digestion, three methods developed from hexadeeyltrimethylammonium bromide (CTAB) or sodium dodecyl sulfate (SDS) method respectively were optimized and compared in this paper. Of the three modified methods used, method Ⅱ produced pure and high-quantity genomic DNA from small amount samples oftransgenie Dendrobium or other plantlets in vitro tissue culture. DNA isolated by method Ⅱ was proved amenable to PCR amplification, restriction digestion and Southern blot analysis of transgenic Dendrobium plantlets.
DendrobiumDNA isolationPolysaccharidesPCRSouthern blot
张妙彬、潘丽晶、范干群、陈继敏、程萍
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珠海市农业科学研究中心,珠海,519070
石斛兰 DNA提取 多糖 PCR Southern杂交
Science and Technology Plan Project in Guangdong Province of China
NETL
NSTL
维普
万方数据
