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Critical factors for assembling a high volume of DNA barcodes

机译:组装大量DNA条形码的关键因素

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摘要

Large-scale DNA barcoding projects are now moving toward activation while the creation of a comprehensive barcode library for eukaryotes will ultimately require the acquisition of some 100 million barcodes. To satisfy this need, analytical facilities must adopt protocols that can support the rapid, cost-effective assembly of barcodes. In this paper we discuss the prospects for establishing high volume DNA barcoding facilities by evaluating key steps in the analytical chain from specimens to barcodes. Alliances with members of the taxonomic community represent the most effective strategy for provisioning the analytical chain with specimens. The optimal protocols for DNA extraction and subsequent PCR amplification of the barcode region depend strongly on their condition, but production targets of 100K barcode records per year are now feasible for facilities working with compliant specimens. The analysis of museum collections is currently challenging, but PCR cocktails that combine polymerases with repair enzyme(s) promise future success. Barcode analysis is already a cost-effective option for species identification in some situations and this will increasingly be the case as reference libraries are assembled and analytical protocols are simplified.
机译:大型DNA条形码项目现在正朝着激活的方向发展,而为真核生物建立一个全面的条形码库最终将需要获得约1亿个条形码。为了满足此需求,分析设施必须采用能够支持快速,经济高效地组装条形码的协议。在本文中,我们通过评估从样本到条形码的分析链中的关键步骤,讨论了建立大量DNA条形码设施的前景。与生物分类界成员的联盟代表了为分析链提供标本的最有效策略。 DNA提取和条形码区域随后进行PCR扩增的最佳方案在很大程度上取决于其条件,但现在每年处理10万条条形码记录的生产目标对于使用顺应性标本的设施来说是可行的。博物馆藏品的分析目前具有挑战性,但结合聚合酶和修复酶的PCR混合物有望获得成功。在某些情况下,条形码分析已经是一种有效的物种识别方法,并且随着参考库的组装和分析协议的简化,情况将越来越多。

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