一种从PAXgene全血RNA管内提取基因组DNA的方法

摘要

目的:从同一生物样本同步提取RNA和DNA,能提高样本的利用率,而且对于基因组学、转录组学和表观遗传学检测数据之间的比对和匹配分析也十分重要.本研究在不影响RNA样品制备的前提下,建立一种从PAXgene全血RNA管内提取基因组DNA的方法.方法:取一定量PAXgene全血RNA管血液样本,使用QIAamp DNA试剂盒提取血细胞基因组DNA,系统优化提取过程中的离心参数、洗脱量以及初始血液样本量等实验参数,并对提取的基因组DNA质量进行检测.结果:用PAXgene全血RNA管3mL血液样本能够提取出8.918±1.100μg基因组DNA,紫外分光光度计检测DNA样品的OD 260/280比值为1.89±0.09,琼脂糖凝胶电泳结果显示DNA样品完整无降解.结论:利用本方法提取的DNA样品能够满足下游DNA芯片、DNA甲基化测序等实验要求.该方法有助于从有限的临床血液样本中获取全面的遗传信息,并且提高后续不同实验方法所生成数据之间的可比性和匹配度.%Objective:Simultaneous extraction of RNA and DNA from the same biological sample can improve the utilization of the sample,and it is also very important for the comparison and analysis of the data of genomics,proteomics and epigenetic detection.This study established a method for extracting genomic DNA from PAXgene blood RNA tube without affecting the preparation of RNA samples.Methods:A certain amount of blood samples was taken and gDNA was extracted from blood cells using the QIAamp nucleic acid purification kit.The centrifugal parameters including centrifugal speed and time,elution volume and initial blood sample volume were optimized in the process of extraction and then the quality of the extracted gDNA was detected.Results:We could extract 8.918 ± 1.100 μg (mean ± SD) gDNA from 3 mL tube-sample with an OD 260/280 ratio of 1.89± 0.09 (mean ± SD).The agarose gel electrophoresis analysis showed that the extracted gDNA was of high molecular weight and integrity.Conclusion:The extracted gDNA is generally able to meet the requirements of downstream experiments such as DNA chip,DNA methylation sequencing.This method is helpful to obtain comprehensive genetic information from the limited clinical blood samples,and improve the comparability and the matching degree between the data generated by different experimental methods.