不同产地连翘新鲜果实DNA提取方法和质量评价研究

目的建立连翘新鲜果实中高质量DNA的提取方法,为从分子水平研究和评价连翘药材质量奠定基础。方法采用改良的CTAB法提取连翘的总DNA,应用琼脂糖凝胶电泳法、分光光度法与PCR法对连翘DNA产率和质量进行检测评价。结果 14批连翘药材核酸蛋白仪测定值A260/A280均在1.7~1.9之间,改良CTAB法提取的连翘总DNA浓度较大,干扰较小,较高质量的DNA可进行PCR扩增,1%琼脂糖凝胶电泳之后用紫外成像仪观察结果正常。结论改良CTAB法适合连翘新鲜果实中DNA的提取,方法简单可靠。

Study on DNA Extraction Method and Quality Evaluation of Forsythia Fresh Fruit of Different Places

Objective ToestablishtheextractionmethodofhighqualityDNAinforsythiafreshfruit so as to lay the foundation for molecular research and evaluation of forsythia medical material quality.Meth-ods ThemodifiedCTABmethodwasadoptedtoextractthetotalDNAofforsythia.Theagarosegelelectro-phoresis（AGE）,spectrophotometry and PCR method were used to evaluate forsythia DNA production and quality.Results A260/A280valueofproteinnucleicacidanalyzerwas1.7to1.9forall14batchesoffor-sythia.The concentration of total DNA extracted with modified CTAB was higher and the interference was less.PCR proliferation was applied to DNA of higher quality,and the result was normal under the observation withultravioletimagerafter1%AGE.Conclusion ThemodifiedCTABisapplicableforDNAextractionof forsythia fresh fruit.This method is simple and reliable.