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目的探索适用于Pythiumsp.GY1938菌株简并PCR的基因组DNA模板制备方案。方法采用微波法、石英沙法、氯化苄法、CTAB法、尿素法提取DNA,并通过特异PCR和简并PCR评价各种方案的优弊。结果 5种方法制备的DNA含量差异无统计学意义(F=2.355,P>0.05);DNA制备效率相近;特异性PCR结果一致,但氯化苄法制备的DNA经简并PCR扩增出的条带较多,目的条带更清晰。结论 5种方法均适于特异性PCR所需DNA模板的制备,但氯化苄法更适于Pythiumsp.GY1938菌株简并PCR模板DNA的制备。
Objective To explore a genomic DNA template preparation method suitable for degenerative PCR of Pythium sp. GY1938 strain. Methods Microwave, quartz sand, benzyl chloride, CTAB and urea were used to extract DNA. The advantages and disadvantages of different protocols were evaluated by specific PCR and degenerate PCR. Results There was no significant difference in DNA content between the five methods (F = 2.355, P> 0.05). The efficiency of DNA preparation was similar and the results of specific PCR were consistent. However, DNA prepared by benzyl chloride method was amplified by degenerate PCR More strips, the purpose of the strip clearer. Conclusion All five methods are suitable for the preparation of DNA template for specific PCR, but the benzyl chloride method is more suitable for the preparation of degenerate PCR template DNA of Pythium sp. GY1938 strain.