Improving PacBio long read accuracy by short read alignment.

Improving PacBio long read accuracy by short read alignment.
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DOI:
10.1371/journal.pone.0046679
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发表时间:
2012
期刊:
影响因子:
3.7
通讯作者:
Wong WH
Wong WH
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Au KF;Underwood JG;Lee L;Wong WH

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最近发展的第三代测序 (TGS) 产生的读数比第二代测序 (SGS) 长得多,从而为解决仅通过 SGS 难以研究的问题提供了机会。然而,较高的原始读取错误率是大多数 TGS 技术的固有缺点。在这里,我们提出了一种计算方法 LSC,通过 SGS 短读取 (SR) 对 TGS 长读取 (LR) 进行纠错。为了降低主 TGS 平台中均聚物运行的错误率,PacBio® RS、LSC 采用均聚物压缩 (HC) 转换策略来提高 SR-LR 比对的灵敏度,而不影响比对精度。我们将 LSC 应用于人脑小脑 RNA-seq 数据中的 100,000 个 PacBio 长读数和人脑 RNA-seq 数据中的 6400 万个单端 75 bp 读数。结果显示LSC可以纠正PacBio长读,将错误率降低3倍以上。准确性的提高极大地有利于许多下游分析,例如 RNA-seq 研究中的定向基因亚型检测。与另一种混合校正工具相比,LSC 可以实现双倍以上的灵敏度和相似的特异性。
The recent development of third generation sequencing (TGS) generates much longer reads than second generation sequencing (SGS) and thus provides a chance to solve problems that are difficult to study through SGS alone. However, higher raw read error rates are an intrinsic drawback in most TGS technologies. Here we present a computational method, LSC, to perform error correction of TGS long reads (LR) by SGS short reads (SR). Aiming to reduce the error rate in homopolymer runs in the main TGS platform, the PacBio® RS, LSC applies a homopolymer compression (HC) transformation strategy to increase the sensitivity of SR-LR alignment without scarifying alignment accuracy. We applied LSC to 100,000 PacBio long reads from human brain cerebellum RNA-seq data and 64 million single-end 75 bp reads from human brain RNA-seq data. The results show LSC can correct PacBio long reads to reduce the error rate by more than 3 folds. The improved accuracy greatly benefits many downstream analyses, such as directional gene isoform detection in RNA-seq study. Compared with another hybrid correction tool, LSC can achieve over double the sensitivity and similar specificity.
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