A cost-effective method for high-throughput construction of illumina sequencing libraries.
A cost-effective method for high-throughput construction of illumina sequencing libraries.
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DOI：
10.1101/pdb.prot074187
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发表时间：
2013-09-01
影响因子：
--
通讯作者：
Friesen ML
中科院分区：
文献类型：
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作者：
Dunham JP;Friesen ML
尽管下一代测序的成本急剧下降，但使用商用试剂盒制备测序文库仍然很昂贵。对于需要分析数百到数千个样本的大规模比较或实验研究来说，成本可能令人望而却步。越来越多地使用数十到数百个样品的多路复用强调了迫切需要开发一种具有成本效益和时间效率的高通量文库制备方法。通过优化和缩小文库建设的步骤，并使用常用的试剂，这里描述的协议允许在96孔格式下制备DNA文库，不需要专门的设备，并且在试剂成本和人员时间上都有很大的节省。与市售试剂盒相比，使用这种优化的高通量格式可将成本降低10倍，单个制备库的每个库或池样成本为~ 12.60-14.90美元，单个条形码池库的成本为~ 8.60-10.60美元；这两种技术允许多达144个样品汇集在一个单车道与条形码测试在这里。
Despite the plummeting cost of next-generation sequencing, the preparation of sequencing libraries using commercially available kits still remains expensive. The cost can be prohibitive for large-scale comparative or experimental studies, where hundreds to thousands of samples need to be analyzed. The increasing use of multiplexing dozens to hundreds of samples underscores the urgent need to develop a cost-effective and time-efficient high-throughput method for library preparation. By optimizing and scaling down the steps in library construction and using commonly available reagents, the protocol described here allows for the preparation of DNA libraries in a 96-well format using no specialized equipment and at a substantial savings in both reagent cost and personnel hours. Utilizing this optimized high-throughput format results in a 10-fold cost reduction, compared to commercially available kits, making per library or pooled sample costs∼ $12.60–14.90 for individually prepared libraries and∼ $8.60–10.60 for pooled libraries with individual barcodes; both techniques allow for up to 144 samples to be pooled on a single lane with the barcodes tested herein.