Highly-multiplexed barcode sequencing: an efficient method for parallel analysis of pooled samples.
Highly-multiplexed barcode sequencing: an efficient method for parallel analysis of pooled samples.
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DOI：
10.1093/nar/gkq368
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发表时间：
2010-07
影响因子：
14.9
通讯作者：
Nislow C
中科院分区：
文献类型：
--
作者：
Smith AM;Heisler LE;St Onge RP;Farias-Hesson E;Wallace IM;Bodeau J;Harris AN;Perry KM;Giaever G;Pourmand N;Nislow C
下一代测序已被证明是分子计数应用的一种极其有效的技术，其中序列读数为RNA-SEQ、CHIP-SEQ、TN-SEQ和其他应用提供数字读数。每次运行可获得的极大数量的序列读数允许分析日益复杂的样品。然而，对于复杂度较低的样本，当每个序列的计数数量导致过采样而不增加数据质量时，达到了回报递减的点。使下一代测序尽可能高效和负担得起的解决方案包括在一次运行中分析多个样本。在这里，我们报告了对DNA条码酵母突变复合库的成功的96-plex，并表明对这些样本的这种‘Bar-Seq’评估可以与条码微阵列提供的数据相媲美，条码微阵列是这一应用的当前基准。高度多重测序所允许的成本降低和吞吐量的增加将极大地扩展化学基因组分析的范围，同样重要的是，该方法适用于其他可能受益于大规模并行化的序列计数应用。
Next-generation sequencing has proven an extremely effective technology for molecular counting applications where the number of sequence reads provides a digital readout for RNA-seq, ChIP-seq, Tn-seq and other applications. The extremely large number of sequence reads that can be obtained per run permits the analysis of increasingly complex samples. For lower complexity samples, however, a point of diminishing returns is reached when the number of counts per sequence results in oversampling with no increase in data quality. A solution to making next-generation sequencing as efficient and affordable as possible involves assaying multiple samples in a single run. Here, we report the successful 96-plexing of complex pools of DNA barcoded yeast mutants and show that such ‘Bar-seq’ assessment of these samples is comparable with data provided by barcode microarrays, the current benchmark for this application. The cost reduction and increased throughput permitted by highly multiplexed sequencing will greatly expand the scope of chemogenomics assays and, equally importantly, the approach is suitable for other sequence counting applications that could benefit from massive parallelization.
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影响因子：
4.5
作者：
Ericson E;Gebbia M;Heisler LE;Wildenhain J;Tyers M;Giaever G;Nislow C
通讯作者：
Nislow C
影响因子：
30.8
作者：
Alkan, Can;Kidd, Jeffrey M.;Marques-Bonet, Tomas;Aksay, Gozde;Antonacci, Francesca;Hormozdiari, Fereydoun;Kitzman, Jacob O.;Baker, Carl;Malig, Maika;Mutlu, Onur;Sahinalp, S. Cenk;Gibbs, Richard A.;Eichler, Evan E.
通讯作者：
Eichler, Evan E.
影响因子：
46.9
作者：
通讯作者：
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影响因子：
48
作者：
Craig, David W.;Pearson, John V.;Szelinger, Szabolcs;Sekar, Aswin;Redman, Margot;Corneveaux, Jason J.;Pawlowski, Traci L.;Laub, Trisha;Nunn, Gary;Stephan, Dietrich A.;Homer, Nils;Huentelman, Matthew J.
通讯作者：
Huentelman, Matthew J.
影响因子：
56.9
作者：
Hillenmeyer, Maureen E.;Fung, Eula;Giaever, Guri
通讯作者：
Giaever, Guri