DNA damage caused by common cytological fixatives
DNA damage caused by common cytological fixatives
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DOI：
10.1016/s0027-5107(97)00314-x
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发表时间：
1998-06-05
影响因子：
2.3
通讯作者：
Rogers, SO
中科院分区：
文献类型：
--
作者：
Douglas, MP;Rogers, SO
分别用7种细胞学固定剂(3.7%甲醛在pH 3.0和7.0，FAA在pH 3.0和7.0，1%戊二醛在pH 3.0和7，1%戊二醛在pH 3.0和7.0，Lavdowsky液在pH 3.0)和1种储存缓冲液(SED=NaCl-EDTA-DMSO，pH 7.0)分别处理9种植物和真菌的组织。未经处理的组织和经SED处理的组织的DNA具有高分子量(>；50kb)。来自戊二醛处理组织的DNA平均长度为20kb，而来自所有其他处理的DNA平均长度不到8kb。用聚合酶链式反应对每个DNA进行扩增，然后对核rRNA小亚基基因3‘端附近的250bp进行测序。戊二醛处理(pH 3.0和7.0)产生的碱基损伤率为0.0%到不到0.1%。用拉夫多斯基液(含氯化汞)、pH7.0的FAA和SED治疗，可产生0.0%至3.6%的比率。在pH值为3.0时，FAA的产率为7.6%~15.6%。用3.7%的甲醛(在pH 3.0和7.0)处理的标本进行了近100次扩增，但都失败了，这表明DNA受到了极大的破坏。(C)1998 Elsevier Science B.V.保留所有权利。
Tissues from nine species of plants and fungi were treated separately with eight solutions, including seven cytological fixatives (3.7% formaldehyde at pH 3.0 and 7.0, FAA at pH 3.0 and 7.0, 1% glutaraldehyde at pH 3.0 and 7,0, and Lavdowsky's fluid at pH 3.0) and one storage buffer (SED = NaCl-EDTA-DMSO, pH 7.0). DNA from untreated tissue and SED-treated tissue was of high molecular weight (> 50 kb). DNA from glutaraldehyde-treated tissues averaged 20 kb in length, while DNA from all other treatments averaged less than 8 kb in length. Each DNA was subjected to amplification using the polymerase chain reaction, followed by sequencing of 250 bp near the 3' end of the nuclear rRNA small subunit gene. Glutaraldehyde treatments (at pH 3.0 and 7.0) produced damaged bases at rates of 0.0% to less than 0.1%. Treatments with Lavdowsky's fluid (containing mercuric chloride), FAA at pH 7.0, and SED produced rates of 0.0% to 3.6%. FAA at pH 3.0 produced rates of 7.6% to 15.6%. Nearly 100 attempts to amplify from specimens treated with 3.7% formaldehyde (at pH 3.0 and 7.0) failed, indicating extreme damage to the DNA. (C) 1998 Elsevier Science B.V. All rights reserved.