School of Chemistry, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
Physical Sciences of Imaging in the Biomedical Sciences Centre for Doctoral Training, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
Nucleic Acids Res. 2019 Jul 9;47(12):e68. doi: 10.1093/nar/gkz212.
We report an approach for visualizing DNA sequence and using these 'DNA barcodes' to search complex mixtures of genomic material for DNA molecules of interest. We demonstrate three applications of this methodology; identifying specific molecules of interest from a dataset containing gigabasepairs of genome; identification of a bacterium from such a dataset and, finally, by locating infecting virus molecules in a background of human genomic material. As a result of the dense fluorescent labelling of the DNA, individual barcodes of the order 40 kb pairs in length can be reliably identified. This means DNA can be prepared for imaging using standard handling and purification techniques. The recorded dataset provides stable physical and electronic records of the total genomic content of a sample that can be readily searched for a molecule or region of interest.
我们报告了一种可视化 DNA 序列的方法,并利用这些“DNA 条码”从基因组物质的复杂混合物中搜索感兴趣的 DNA 分子。我们展示了该方法的三种应用;从包含千兆碱基对基因组的数据集识别特定的感兴趣分子;从这样的数据集识别细菌,最后,通过定位感染病毒分子在人类基因组物质的背景中。由于 DNA 的密集荧光标记,可以可靠地识别长度约为 40kb 对的单个条码。这意味着可以使用标准的处理和纯化技术来准备用于成像的 DNA。记录的数据集为样品的总基因组内容提供了稳定的物理和电子记录,可以方便地搜索感兴趣的分子或区域。