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小鼠细菌人工染色体末端质量评估及序列分析。

Mouse BAC ends quality assessment and sequence analyses.

作者信息

Zhao S, Shatsman S, Ayodeji B, Geer K, Tsegaye G, Krol M, Gebregeorgis E, Shvartsbeyn A, Russell D, Overton L, Jiang L, Dimitrov G, Tran K, Shetty J, Malek J A, Feldblyum T, Nierman W C, Fraser C M

机构信息

The Institute for Genomic Research, Rockville, Maryland 20850, USA.

出版信息

Genome Res. 2001 Oct;11(10):1736-45. doi: 10.1101/gr.179201.

Abstract

A large-scale BAC end-sequencing project at The Institute for Genomic Research (TIGR) has generated one of the most extensive sets of sequence markers for the mouse genome to date. With a sequencing success rate of >80%, an average read length of 485 bp, and ABI3700 capillary sequencers, we have generated 449,234 nonredundant mouse BAC end sequences (mBESs) with 218 Mb total from 257,318 clones from libraries RPCI-23 and RPCI-24, representing 15x clone coverage, 7% sequence coverage, and a marker every 7 kb across the genome. A total of 191,916 BACs have sequences from both ends providing 12x genome coverage. The average Q20 length is 406 bp and 84% of the bases have phred quality scores > or = 20. RPCI-24 mBESs have more Q20 bases and longer reads on average than RPCI-23 sequences. ABI3700 sequencers and the sample tracking system ensure that > 95% of mBESs are associated with the right clone identifiers. We have found that a significant fraction of mBESs contains L1 repeats and approximately 48% of the clones have both ends with > or = 100 bp contiguous unique Q20 bases. About 3% mBESs match ESTs and > 70% of matches were conserved between the mouse and the human or the rat. Approximately 0.1% mBESs contain STSs. About 0.2% mBESs match human finished sequences and > 70% of these sequences have EST hits. The analyses indicate that our high-quality mouse BAC end sequences will be a valuable resource to the community.

摘要

美国基因组研究所(TIGR)开展的一项大规模细菌人工染色体(BAC)末端测序项目,已生成了迄今为止最为全面的小鼠基因组序列标记集之一。凭借超过80%的测序成功率、平均485碱基对的读长以及ABI3700毛细管测序仪,我们从RPCI - 23和RPCI - 24文库的257,318个克隆中生成了449,234条非冗余小鼠BAC末端序列(mBESs),总长度达218兆碱基,覆盖了15倍的克隆覆盖率、7%的序列覆盖率,且全基因组平均每7千碱基就有一个标记。共有191,916个BAC两端都有序列,提供了12倍的基因组覆盖率。平均Q20长度为406碱基对,84%的碱基的Phred质量得分大于或等于20。RPCI - 24的mBESs平均比RPCI - 23序列有更多Q20碱基和更长的读长。ABI3700测序仪和样本追踪系统确保超过95%的mBESs与正确的克隆标识符相关联。我们发现相当一部分mBESs包含L1重复序列,约48%的克隆两端都有大于或等于100碱基对的连续唯一Q20碱基。约3%的mBESs与表达序列标签(ESTs)匹配,且超过70%的匹配在小鼠与人或大鼠之间是保守的。约0.1%的mBESs包含序列标签位点(STSs)。约0.2%的mBESs与人类完成序列匹配,其中超过70%的这些序列有EST匹配。分析表明,我们高质量的小鼠BAC末端序列将成为该领域的宝贵资源。

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