Best Practices in Designing, Sequencing, and Identifying Random DNA Barcodes.

Best Practices in Designing, Sequencing, and Identifying Random DNA Barcodes.
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DOI:
10.1007/s00239-022-10083-z
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发表时间:
2023-06
影响因子:
3.9
通讯作者:
Kryazhimskiy, Sergey
Kryazhimskiy, Sergey
中科院分区:
生物学3区
文献类型:
--
作者:
Johnson, Milo S.;Venkataram, Sandeep;Kryazhimskiy, Sergey

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随机DNA条形码是一种跟踪细胞谱系的通用工具,应用范围从发育到癌症再到进化。在这里,我们回顾并严格评估条形码设计以及条形码排序和条形码数据的初始处理的方法。我们首先演示各种条形码设计决策如何影响数据质量,并提出一种新的设计,它平衡了我们目前所知道的所有考虑因素。然后,我们讨论了制备条形码测序库的各种选择,包括行内索引和唯一分子识别符(UMI)。最后,我们测试了几个已建立的和新的生物信息管道的性能,这些管道用于从原始测序读数中提取条形码并进行纠错。我们发现,对齐和基于正则表达式的方法都能很好地用于条形码提取,并且专门为条形码数据设计的纠错管道优于通用管道。总体而言,这篇综述将帮助研究人员以一种深思熟虑和系统的方式处理他们的条形码实验。网上版载有补充材料,可在10.1007/s00239-022-10083-z查阅。
Random DNA barcodes are a versatile tool for tracking cell lineages, with applications ranging from development to cancer to evolution. Here, we review and critically evaluate barcode designs as well as methods of barcode sequencing and initial processing of barcode data. We first demonstrate how various barcode design decisions affect data quality and propose a new design that balances all considerations that we are currently aware of. We then discuss various options for the preparation of barcode sequencing libraries, including inline indices and Unique Molecular Identifiers (UMIs). Finally, we test the performance of several established and new bioinformatic pipelines for the extraction of barcodes from raw sequencing reads and for error correction. We find that both alignment and regular expression-based approaches work well for barcode extraction, and that error-correction pipelines designed specifically for barcode data are superior to generic ones. Overall, this review will help researchers to approach their barcoding experiments in a deliberate and systematic way. The online version contains supplementary material available at 10.1007/s00239-022-10083-z.
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