EXTRACTION OF DNA FROM SHEEP WHITE BLOOD-CELLS
EXTRACTION OF DNA FROM SHEEP WHITE BLOOD-CELLS
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DOI：
10.1080/00288233.1990.10428440
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发表时间：
1990-01-01
影响因子：
1.8
通讯作者：
SISE, JA
中科院分区：
文献类型：
--
作者：
MONTGOMERY, GW;SISE, JA
从全血中提取DNA的常用方法包括几个酚提取步骤。苯酚具有腐蚀性和毒性，提取步骤耗时，限制了可以处理的样品数量。用高盐法和盐酸胍法两种方法从羊血中提取DNA。使用盐酸胍方法提取得到凝胶状物质，其无法重悬于TE缓冲液中。琼脂糖凝胶电泳测定，高盐方法产生了良好的高分子量DNA产率。用酚法和高盐法提取的100份20 ml全血DNA的平均得率分别为0.50 mg（SD = 0.19）和0.64 mg（SD = 0.26）。高盐法提取的DNA质量与苯酚法相当，两种方法提取的DNA均能用多种限制性内切酶完全消化，适合Southern印迹分析。高盐方法可常规用于从绵羊样品中提取DNA，增加处理的样品数量，并促进DNA提取用于遗传连锁分析和诊断测试。
Common methods for the extraction of DNA from whole blood involve several phenol extraction steps. Phenol is corrosive and toxic, and the extraction steps aretime-consuming, limiting the number of samples that can be processed. Two other methods using high salt or guanidine hydrochloride were tested for extraction of DNA from sheep blood. Extraction using the guanidine hydrochloride method resulted in a gelatinous material that failed to resuspend in TE buffer. The high salt method produced a good yield of high molecular weight DNA as determined agarose gel electrophoresis. The mean yields of DNA from 100 samples of 20 ml of whole blood extracted by either the phenol or the high salt methods were 0.50 mg (SD = 0.19) and 0.64 mg (SD = 0.26) respectively. The quality of the DNA extracted by th high salt method was equivalent to the extracted by the phenol method and the DNA from both methods was digested to completion with a range of restriction enzymes and was suitable for analysis by Southern blotting. The high salt method can be used routinely for the extraction of DNA from sheep samples, increasing the number of samples processed and facilitating DNA extraction for genetic linkage analysis and diagnostic tests.