中国组织工程研究2012，Vol.16Issue(11)：1973-1976,4.DOI:10.3969/j.issn.1673-8225.2012.11.017
石蜡包埋组织提取DNA不同条件下的PCR扩增
PCR amplication of the DNA extracted from paraffin-embedded tissues in different conditions
马丽丽 1艾力江·吐尔逊 1张莉1
作者信息
- 1. 新疆医科大学第一附属医院肿瘤中心,新疆维吾尔自治区,乌鲁木齐市,830011
- 折叠
摘要
Abstract
BACKGROUND: Because of the damage of formalin-fixed paraffin-embedded tissue to DNA in the process of its production andpreservation, it is difficult to extract high quality and sufficient DNA for PCR.OBJECTIVE: To analyze the influencing factors of the DNA extracted from paraffin-embedded tissue for PCR.METHODS: Genomic DNA was extracted from paraffin-embedded esophageal carcinoma tissue sections of 10.0 μm×2,10.0 μm×4and 10.0 μm×5. Template DNA amount of 0.05, 0.1, 0.2 μg was used to amplify gene β-actin, respectively. PCR cycletimes were set as 35, 40 and 45. The influencing factors of PCR were analyzed.RESULTS AND CONCLUSION: The analysis of agarose gel electrophoresis showed that when paraffin-embedded esophagealcarcinoma tissue section amount was 10.0 μm×2, the PCR cycle times were 40; when the template DNA amount was 0.05μg, theobtained target DNA was of the highest quality. It shows that reducing the amount of paraffin-embedded tissue and DNA templatecan be help to require high quality PCR strips; PCR cycle times should be more than 40 and less than 45, and if it increasesbeyond this range there is meaningless.关键词
石蜡包埋组织/提取DNA/PCR/反应/循环次数/模板DNA分类
医药卫生引用本文复制引用
马丽丽,艾力江·吐尔逊,张莉..石蜡包埋组织提取DNA不同条件下的PCR扩增[J].中国组织工程研究,2012,16(11):1973-1976,4.
