Optimized Replication of Arrayed Bacterial Mutant Libraries Increases Access to Biological Resources.
Optimized Replication of Arrayed Bacterial Mutant Libraries Increases Access to Biological Resources.
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DOI：
10.1128/spectrum.01693-23
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发表时间：
2023-08-17
影响因子：
3.7
通讯作者：
Dunny, Gary M. M.
中科院分区：
文献类型：
--
作者：
Willett, Julia L. E.;Barnes, Aaron M. T.;Brunson, Debra N. N.;Lecomte, Alexandre;Robertson, Ethan B. B.;Dunny, Gary M. M.
关键词：
生物样本库，包括单转座子（Tn）或缺失突变体的阵列文库，极大地加快了细菌遗传学研究的步伐。尽管这些资源很重要，但关于这些材料的复制和分发的操作流程却很少。在此，我们描述了一种用于创建一个阵列细菌Tn文库多个复制品的操作流程，该文库在96孔板（73个板）中包含约6800个突变体。我们的操作流程提供了多个检查点，以防止污染并将冻融循环导致的遗传漂变降至最低。这种方法也可适用于各种规模的阵列培养物样本库。总体而言，对于考虑构建和分发阵列培养物样本库资源以造福更广泛的科学界的其他研究人员来说，该操作流程是一种宝贵的资源。
重要性：阵列突变体样本库推动了稳健的遗传筛选，但关于这些资源的复制以及后续质量控制的操作流程却很少。增加阵列生物样本库的分发将提高这些资源的可获取性和利用率。开发用于复制这些资源的标准化技术对于确保其质量以及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. IMPORTANCE Arrayed 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.
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影响因子：
4.5
作者：
Chatterjee A;Willett JLE;Dunny GM;Duerkop BA
通讯作者：
Duerkop BA
影响因子：
6.4
作者：
Dale JL;Beckman KB;Willett JLE;Nilson JL;Palani NP;Baller JA;Hauge A;Gohl DM;Erickson R;Manias DA;Sadowsky MJ;Dunny GM
通讯作者：
Dunny GM
影响因子：
6.4
作者：
Fey PD;Endres JL;Yajjala VK;Widhelm TJ;Boissy RJ;Bose JL;Bayles KW
通讯作者：
Bayles KW
影响因子：
3.7
作者：
Ortlieb N;Klenk E;Kulik A;Niedermeyer THJ
通讯作者：
Niedermeyer THJ
影响因子：
30.3
作者：
Keogh D;Tay WH;Ho YY;Dale JL;Chen S;Umashankar S;Williams RBH;Chen SL;Dunny GM;Kline KA
通讯作者：
Kline KA