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一种从PAXgene全血RNA管内提取基因组DNA的方法
  • ISSN号:1673-6273
  • 期刊名称:《现代生物医学进展》
  • 时间:0
  • 分类:Q751[生物学—分子生物学] R-331[医药卫生]
  • 作者机构:上海中医药大学上海市针灸经络研究所,上海200030
  • 相关基金:国家自然科学基金项目(81574058,81473760);上海市中医药事业发展三年行动计划项目(ZY3-CCCX-3-3005,ZYSNXD-CC-ZDYJ039);上海市卫生和计划生育委员会科研课题(201640133)
中文摘要:

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3 m L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。

英文摘要:

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.

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期刊信息
  • 《现代生物医学进展》
  • 中国科技核心期刊
  • 主管单位:黑龙江省卫生厅
  • 主办单位:黑龙江省红十字医院 黑黑龙江省红十字医院 黑龙江省森林工总医院
  • 主编:申宝忠
  • 地址:哈尔滨市南岗区花园街184号403
  • 邮编:150001
  • 邮箱:biomed_54@126.com
  • 电话:0451-82583800 53658268
  • 国际标准刊号:ISSN:1673-6273
  • 国内统一刊号:ISSN:23-1544/R
  • 邮发代号:14-12
  • 获奖情况:
  • 国内外数据库收录:
  • 被引量:33230