Department of Microbiology, University of Pennsylvania, Philadelphia, PA, USA.
Department of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Bioinformatics. 2017 Jul 15;33(14):2071-2077. doi: 10.1093/bioinformatics/btx118.
Population genomic analyses are often hindered by difficulties in obtaining sufficient numbers of genomes for analysis by DNA sequencing. Selective whole-genome amplification (SWGA) provides an efficient approach to amplify microbial genomes from complex backgrounds for sequence acquisition. However, the process of designing sets of primers for this method has many degrees of freedom and would benefit from an automated process to evaluate the vast number of potential primer sets.
Here, we present swga , a program that identifies primer sets for SWGA and evaluates them for efficiency and selectivity. We used swga to design and test primer sets for the selective amplification of Wolbachia pipientis genomic DNA from infected Drosophila melanogaster and Mycobacterium tuberculosis from human blood. We identify primer sets that successfully amplify each against their backgrounds and describe a general method for using swga for arbitrary targets. In addition, we describe characteristics of primer sets that correlate with successful amplification, and present guidelines for implementation of SWGA to detect new targets.
Source code and documentation are freely available on https://www.github.com/eclarke/swga . The program is implemented in Python and C and licensed under the GNU Public License.
Supplementary data are available at Bioinformatics online.
群体基因组分析通常受到通过 DNA 测序获得足够数量的基因组进行分析的困难的阻碍。选择性全基因组扩增(SWGA)为从复杂背景中扩增微生物基因组以进行序列获取提供了一种有效的方法。然而,为该方法设计引物集的过程具有许多自由度,并且从自动评估大量潜在引物集的过程中受益。
在这里,我们提出了 swga,这是一个用于识别 SWGA 引物集并评估其效率和选择性的程序。我们使用 swga 设计并测试了从感染的果蝇和人类血液中的结核分枝杆菌中选择性扩增沃尔巴克氏体 pipientis 基因组 DNA 的引物集。我们确定了针对各自背景成功扩增的引物集,并描述了一种使用 swga 检测任意目标的通用方法。此外,我们描述了与成功扩增相关的引物集的特征,并提出了用于实施 SWGA 以检测新目标的指南。
源代码和文档可在 https://www.github.com/eclarke/swga 上免费获得。该程序是用 Python 和 C 编写的,并根据 GNU 公共许可证获得许可。
补充数据可在生物信息学在线获得。