Optimized replication of arrayed bacterial mutant libraries increase access to biological resources.
Optimized replication of arrayed bacterial mutant libraries increase access to biological resources.
复制标题
阵列细菌突变体文库的优化复制增加了生物资源的获取。
DOI：
10.1101/2023.04.25.537918
复制
发表时间：
2023
期刊：
影响因子：
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通讯作者：
Dunny,GaryM
中科院分区：
文献类型：
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作者：
Willett,JuliaLE;Barnes,AaronMT;Brunson,DebraN;Lecomte,Alexandre;Robertson,EthanB;Dunny,GaryM
生物集合，包括单个转座子（Tn）或缺失突变体的排列文库，极大地加快了细菌遗传研究的步伐。尽管这些资源很重要，但很少有复制和分发这些材料的协议。在这里，我们描述了一种在96孔板（73板）中创建由大约6800个突变体组成的排列细菌Tn文库的多个复制的方案。我们的方案提供了多个检查点，以防止污染，并尽量减少由冻融循环引起的遗传漂变。此方法还可以针对各种大小的已排列区域性集合进行缩放。总的来说，该协议对于其他考虑构建和分配阵列培养收集资源的研究人员来说是一个有价值的资源，有利于更大的科学界。重要的是，测序的突变体集合驱动强大的遗传筛选，但很少有协议存在这些资源的复制和随后的质量控制。增加排列生物馆藏的分布将增加这些资源的可及性和利用率。开发复制这些资源的标准化技术对于确保它们的质量和对科学界的有用性至关重要。
Biological collections, including arrayed libraries of single transposon (Tn) or deletion mutants, greatly accelerate the pace of bacterial genetic research. Despite the importance of these resources, few protocols exist for the replication and distribution of these materials. Here, we describe a protocol for creating multiple replicates of an arrayed bacterial Tn library consisting of approximately 6,800 mutants in 96-well plates (73 plates). Our protocol provides multiple checkpoints to guard against contamination and minimize genetic drift caused by freeze/thaw cycles. This approach can also be scaled for arrayed culture collections of various sizes. Overall, this protocol is a valuable resource for other researchers considering the construction and distribution of arrayed culture collection resources for the benefit of the greater scientific community.IMPORTANCEArrayed mutant collections drive robust genetic screens, but few protocols exist for replication of these resources and subsequent quality control. Increasing the distribution of arrayed biological collections will increase the accessibility and use of these resources. Developing standardized techniques for replication of these resources is essential for ensuring their quality and usefulness to the scientific community.