• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biopreservation and biobanking >Preanalytical Procedures for DNA Studies: The Experience of the Interinstitutional Multidisciplinary BioBank (BioBIM)
【24h】

Preanalytical Procedures for DNA Studies: The Experience of the Interinstitutional Multidisciplinary BioBank (BioBIM)

机译：DNA研究的分析程序：跨机构跨学科生物银行（BioBIM）的经验

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Preanalytical variables, including the anticoagulants and stabilizing agents, time, storage temperature, and methods of DNA extraction applied to blood samples, may affect quality and quantity of isolated nucleic acids for future genomic applications. Considering the large number of collected samples, standard operating procedure optimization for whole blood preservation before DNA extraction is a crucial step in a biological repository. Moreover, the future application of the biological material may not be known subsequent to its extraction. To define standard operating procedures for whole blood preservation before DNA extraction, we aimed to determine whether different combinations of anticoagulants, blood storage temperatures, and time intervals before storage at -80 degrees C might have an impact on quality and quantity of subsequent extracted DNA. After spectrophotometer quantification, the quality and integrity of DNA were assessed by agarose gel electrophoresis, polymerase chain reaction, and real-time polymerase chain reaction methods. We observed that decrease in DNA recovery during blood storage time was more pronounced at room temperature than at 4 degrees C. Based on our experience, we recommend as anticoagulants of choice sodium citrate and ethylenediaminetetraacetate, whereas sodium citrate theophylline adenosine dipyridamole could represent an alternative choice, excluding a priori lithium heparin and Fluoride-Oxalate. Based on the overall evaluation criteria, we conclude that the procedures necessary to preserve the whole blood before the DNA extraction may have a significant impact on downstream molecular biological applications.
机译：分析前的变量，包括抗凝剂和稳定剂，时间，储存温度以及应用于血液样品的DNA提取方法，可能会影响分离的核酸的质量和数量，以供将来的基因组应用。考虑到收集到的大量样本，在DNA提取之前进行全血保存的标准操作程序优化是生物储存库中的关键步骤。而且，在提取生物材料之后，可能不知道生物材料的未来应用。为了定义DNA提取前全血保存的标准操作程序，我们旨在确定抗凝剂，血液存储温度和在-80℃下存储之前的时间间隔的不同组合是否会影响后续提取DNA的质量和数量。分光光度计定量后，通过琼脂糖凝胶电泳，聚合酶链反应和实时聚合酶链反应方法评估DNA的质量和完整性。我们观察到，在血液储存期间，DNA回收率的降低在室温下比在4摄氏度下更为明显。根据我们的经验，我们建议选择柠檬酸钠和乙二胺四乙酸钠作为抗凝剂，而柠檬酸钠茶碱腺苷双嘧达莫可能是另一种选择，不包括先验肝素锂和草酸氟。基于总体评估标准，我们得出结论，在DNA提取之前保存全血所需的程序可能对下游分子生物学应用产生重大影响。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号