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目的比较不同研磨方式、不同提取方法提取的拉萨蒲公英叶片基因组DNA的效果。方法分别用液氮和提取缓冲液两种研磨方式对拉萨蒲公英叶片进行研磨处理,用5种方法提取拉萨蒲公英叶片基因组DNA,分别用紫外分光光度计法、琼脂糖凝胶电泳、PCR扩增检测DNA的质量。结果用提取液研磨可以充分除去色素、酚类、蛋白质等物质,提取的DNA纯度高,降解少,用液氮研磨提取的DNA杂质多,降解严重;高盐低p H法提取的DNA的A260/A280接近1.8,A260/A230为1.93,Ezup柱式试剂盒法提取的DNA的A260/A280为2.01,A260/A230大于2.0,说明这两种方法提取的DNA的纯度比较高,其余3种方法提取的DNA的纯度都比较低。结论用提取缓冲液研磨提取的DNA的质量高于液氮研磨,并且缓冲液研磨能降低DNA提取成本,操作安全,因此是一种经济适用的方法;Ezup柱式试剂盒法操纵简单,但是价格昂贵,且DNA的得率低,高盐低p H法成本低,但是提取时间比试剂盒长,两种方法提取的DNA的PCR效果相差不大,高盐低p H法适用于需要大批量提取拉萨蒲公英叶片DNA的研究。
Objective To compare the effects of different extraction methods on extraction of genomic DNA from leaves of Lhasa dandelion. Methods Taraxacum mongolicum leaves were ground with liquid nitrogen and extraction buffer respectively. Genomic DNA was extracted from leaves of Lhasa dandelion by five different methods. The results were analyzed by ultraviolet spectrophotometer, agarose gel electrophoresis and PCR amplification The quality of DNA. Results The extraction solution was used to remove pigments, phenols, proteins and other substances. The extracted DNA had high purity and less degradation. The extracted DNA was contaminated with liquid nitrogen and had serious degradation. A260 / A280 was close to 1.8, A260 / A230 was 1.93, A260 / A280 of Ezup column kit was 2.01 and A260 / A230 was more than 2.0, which indicated that DNA extracted by these two methods had higher purity. The other three methods The purity of extracted DNA is relatively low. Conclusion The quality of DNA extracted with extraction buffer is higher than that with liquid nitrogen, and buffer grinding can reduce the cost of DNA extraction and safe operation, so it is an economical method. Ezup column method is easy to manipulate, but the price Expensive, low yield of DNA, high salt and low p H method, but the extraction time is longer than the kit, the PCR results of the DNA extracted by the two methods are not much different. The high salt and low p H method is suitable for applications requiring large quantities Study on extraction of DNA from leaves of Lhasa dandelion.