A method for extraction and analysis of high quality genomic DNA from ixodid ticks
A method for extraction and analysis of high quality genomic DNA from ixodid ticks
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DOI：
10.1046/j.1365-2915.2003.00425.x
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发表时间：
2003-06-01
影响因子：
1.9
通讯作者：
Gutierrez, JA
中科院分区：
文献类型：
--
作者：
Hill, CA;Gutierrez, JA
目前，没有公开的方法描述从硬蜱（阿卡里：硬蜱科）中提取高分子量基因组DNA，并且常用的提取方法不能很好地适用于该科的成员。描述了一种从硬蜱中提取最低程度降解的基因组DNA的方法，该方法可以在一天或两天内完成。该方法产生的DNA具有足够的大小（>24 kb）用于Southern分析，并且易于被限制性内切核酸酶消化。Southern分析使用的细胞色素P450基因探针，证明了我们的方法与基因组DNA提取的两种硬蜱，孤独的星星蜱，Amblyomma americanum（Linnaeus）和牛热蜱，微小牛蜱（Canestrini）。
Currently, no published methods describe the extraction of high molecular weight genomic DNA from ixodid ticks (Acari: Ixodidae) and commonly used methods of extraction are not well adapted for use with members of this family. A method for extraction of minimally degraded genomic DNA from ixodid ticks that can be completed in one or two days is described. The method produces DNA which is of sufficient size (>24 kb) for use in Southern analysis and which is readily digestible by restriction endonucleases. Southern analysis using a cytochrome P450 gene probe, demonstrates the success of our method with genomic DNA extracted from two species of Ixodidae, the lone star tick, Amblyomma americanum (Linnaeus) and the cattle fever tick, Boophilus microplus (Canestrini).