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[石蜡包埋材料中的DNA提取及Southern印迹分析]

[DNA extraction and Southern blot analysis in paraffin embedded material].

作者信息

Handt S, Otto F, Wilgenbus K, Kirkpatrick C J

机构信息

Institut für Pathologie der RWTH, Aachen.

出版信息

Verh Dtsch Ges Pathol. 1990;74:378-80.

PMID:1708620
Abstract

DNA was extracted from formaldehyde fixed and paraffin embedded tissue by the use of a modified extraction protocol. In all cases the recovered DNA was more degraded than DNA from fresh or frozen tissue. Fixation times of more than 4 days made it impossible to use the extracted DNA for Southern Blotting; DNA from paraffinized tissues not older than 1 or 2 years could be used for Southern Blotting after digestion with restriction enzymes, but we did not succeed in showing bands with restriction fragments more than 10 kb in length. We conclude that SBA analysis for malignant lymphomas with DNA extracted from routinely paraffin-embedded tissues isn't possible, because the restriction fragments looked for are in many cases longer than 10 kb.

摘要

采用改良的提取方案从甲醛固定石蜡包埋组织中提取DNA。在所有情况下,回收的DNA比新鲜或冷冻组织中的DNA降解程度更高。固定时间超过4天使得无法将提取的DNA用于Southern印迹分析;不超过1或2年的石蜡包埋组织的DNA在用限制性内切酶消化后可用于Southern印迹分析,但我们未能成功显示长度超过10 kb的限制性片段条带。我们得出结论,用常规石蜡包埋组织提取的DNA对恶性淋巴瘤进行SBA分析是不可能的,因为在许多情况下所寻找的限制性片段长度超过10 kb。

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