稻米深加工产品基因组提取方法的研究及其对PCR的影响

本实验以四种米粉为原料，每个样品做三个梯度：120mg, 800mg, 2000mg，采用改进的经典酚/仿法、CTAB沉淀法以及盐酸胍/氯仿法提取基因组，并通过聚合酶链式反应(PCR)通过扩增内标基因(SPS)来检测提取方法的优劣。结果显示：120mg的样品经三种方法提取的基因组均不能扩增出内标基因；800mg的样品和2000mg的样品只有用CTAB沉淀法提取的基因组(采用相同的膜板量)能全部扩增出内标基因。结论：CTAB沉淀法提取基因组的效果最好，对PCR的抑制现象最少。

Study on the Extracted Methods of Genome in Processed Rice Product and Its Influence on PCR

Four kinds of processed rice products were extracted of genomic DNA by the following three methods: classic phenol/chloroform extraction method, CTAB precipitation extraction method and guanidine hydrochloride extraction method. The total DNA quantity can be extracted from different quantity of each processed rice product and the starting material (g) was 120 mg, 800 g and 2000 mg. The quality and quantity of extracted DNA were assayed by PCR with the primer pair SPS-F/SPS-R. Results indicated that all of extracted genomic DNA by the CTAB precipitation extraction method can be amplified by PCR with exception of 0.12 g starting material. So we concluded that the CTAB precipitation extraction method has the highest efficiency of extracting genomic DNA and has least inhibiting information on PCR.