Department of Obstetrics & Gynecology, Columbia University Irving Medical Center, New York, NY 10032, USA.
Department of Pathology & Cell Biology, Columbia University Medical Center, New York, NY 10032, USA.
Biotechniques. 2020 May;68(5):240-244. doi: 10.2144/btn-2019-0172. Epub 2020 Feb 14.
Conventional genomic DNA (gDNA) extraction methods can take hours to complete, may require fume hoods and represent the most time-consuming step in many gDNA-based molecular assays. We systematically optimized a bead bashing-based (BBB) approach for rapid gDNA extraction without the need for a fume hood. Human tissue specimens (n = 34) subjected to the 12-min BBB method yielded 0.40 ± 0.17 (mean ± SD) μg of gDNA per milligram of tissue, sufficient for many downstream applications, and 3- and 6-min extensions resulted in an additional 0.43 ± 0.23 μg and 0.48 ± 0.43 μg per milligram of tissue, respectively. The BBB method provides a simple and rapid method for gDNA extraction from mammalian tissue that is applicable to time-sensitive clinical applications.
传统的基因组 DNA（gDNA）提取方法可能需要数小时才能完成，可能需要通风橱，并且代表了许多基于 gDNA 的分子检测中最耗时的步骤。我们系统地优化了一种基于珠粒捣碎（BBB）的方法，用于快速提取 gDNA，而无需通风橱。经过 12 分钟 BBB 方法处理的人类组织标本（n=34）每毫克组织产生 0.40±0.17（平均值±标准差）μg 的 gDNA，足以满足许多下游应用，分别延长 3 分钟和 6 分钟可使每毫克组织分别额外产生 0.43±0.23μg 和 0.48±0.43μg 的 gDNA。BBB 方法为从哺乳动物组织中提取 gDNA 提供了一种简单、快速的方法，适用于时间敏感的临床应用。
