Effects of delay in the snap freezing of colorectal cancer tissues on the quality of DNA and RNA.
Effects of delay in the snap freezing of colorectal cancer tissues on the quality of DNA and RNA.
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DOI：
10.3393/jksc.2010.26.5.316
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发表时间：
2010-10
期刊：
影响因子：
--
通讯作者：
Park HS
中科院分区：
文献类型：
--
作者：
Hong SH;Baek HA;Jang KY;Chung MJ;Moon WS;Kang MJ;Lee DG;Park HS
使用生物标本进行基础分子研究的成功很大程度上取决于标本的质量。在这项研究中，我们评估了延迟冷冻时间对结直肠癌患者新鲜冰冻组织中DNA和RNA稳定性的影响。20例结直肠癌切除后分别于10、30、60、90min冻存组织。用琼脂糖凝胶电泳法测定A260/A280的吸光度比。此外，还检测了RNA完整性数(RIN)，以分析RNA完整性。不考虑延迟冷冻时间，所有DNA和RNA样品的A260/A280比值均大于1.9，并且所有DNA样品在琼脂糖凝胶电泳图上均显示一条离散的高分子量条带。10、30、60、90min时RIN分别为7.53±2.04、6.70±1.88、6.47±2.58、4.22±2.34。虽然延迟冷冻对RNA的浓度没有影响，但随着延迟冷冻时间的延长，RNA的完整性会降低。根据RIN的结果，对于需要高质量RNA的研究，我们建议结直肠癌组织的收集应该在10分钟内完成，对于常规的RNA研究，我们建议在30分钟内完成。
The success of basic molecular research using biospecimens strongly depends on the quality of the specimen. In this study, we evaluated the effects of delayed freezing time on the stability of DNA and RNA in fresh frozen tissue from patients with colorectal cancer. Tissues were frozen at 10, 30, 60, and 90 minutes after extirpation of colorectal cancer in 20 cases. Absorbance ratio of 260 to 280 nm (A260/A280) and agarose gel electrophoresis were evaluated. In addition, the RNA integrity number (RIN) was assayed for the analysis of the RNA integrity. Regardless of delayed freezing time, all DNA and RNA samples revealed A260/A280 ratios of more than 1.9, and all DNA samples showed a discrete, high-molecular-weight band on agarose gel electrophoresis. The RINs were 7.53 ± 2.04, 6.70 ± 1.88, 6.47 ± 2.58, and 4.22 ± 2.34 at 10, 30, 60, and 90 minutes, respectively. Though the concentration of RNA was not affected by delayed freezing, the RNA integrity was decreased with increasing delayed freezing time. According to the RIN results, we recommend that the collection of colorectal cancer tissue should be done within 10 minutes for studies requiring RNA of high quality and within 30 minutes for usual RNA studies.