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本研究针对一类虫体膨大,体壁硬化,革质化或木质化的球蚧类蚧虫开展了全基因组提取方法的探讨,特别是在材料的选择和前处理过程两个关键环节进行了新的尝试,并通过目前文献报道的SDS、CTAB、TES、NaCl和TNES法进行了随后的基因组提取,运用1%琼脂糖凝胶电泳、DNA OD值测定和特定基因的PCR扩增对基因组的提取效果进行了检测,结果表明,无论是冷冻保存还是75%酒精固定液-4℃保存的球蚧样本均可用于DNA提取,选择球蚧体壁作为提取材料要比整头虫体效果好,前处理采用在少量提取缓冲液中剪碎样品的方法获得的基因组机率大,纯度高、条带整齐无拖尾,同时也证实CTAB法更适合该类蚧虫基因组的提取。
In this study, a wide range of insect body enlargement, body wall sclerosis, leathery or lignified zea scale insects to carry out the whole genome extraction methods, especially in the material selection and pretreatment process two key links A new attempt was made and the subsequent genomic extraction was carried out by SDS, CTAB, TES, NaCl and TNES methods as reported in the literature. The genomic analysis was performed using 1% agarose gel electrophoresis, DNA OD value determination and PCR amplification of specific genes The results showed that either the samples kept in cryopreservation or 75% alcohol fixative at -4 ℃ could be used for DNA extraction. The selection of the body wall of geranium as the extraction material was better than that of the whole body. The pretreatment using the method of cutting samples in a small amount of extraction buffer has the advantages of high genome probability, high purity and no tails of bands, and also confirms that CTAB method is more suitable for extracting the genome of the scale insects.