Identifying essential genes in Mycobacterium tuberculosis by global phenotypic profiling.
Identifying essential genes in Mycobacterium tuberculosis by global phenotypic profiling.
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DOI：
10.1007/978-1-4939-2398-4_6
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发表时间：
2015-01-01
期刊：
影响因子：
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通讯作者：
Sassetti, Christopher M
中科院分区：
文献类型：
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作者：
Long, Jarukit E;DeJesus, Michael;Sassetti, Christopher M
转座子测序（TnSeq）是一种基于深度测序的新一代方法，用于定量评估在选择压力下复杂突变转座子文库的组成。虽然这种方法可以用于任何可能发生转座子突变的生物体，但本章描述了它在结核分枝杆菌中的应用。更具体地说，包括通过转座子诱变、选择压力设计、基因组DNA提取、通过下一代深度测序扩增和定量转座子插入来生成复杂文库的方法。还讨论了基因重要性的确定和收集数据的统计分析。
Transposon sequencing (TnSeq) is a next-generation deep sequencing-based method to quantitatively assess the composition of complex mutant transposon libraries after pressure from selection. Although this method can be used for any organism in which transposon mutagenesis is possible, this chapter describes its use in Mycobacterium tuberculosis. More specifically, the methods for generating complex libraries through transposon mutagenesis, design of selective pressure, extraction of genomic DNA, amplification and quantification of transposon insertions through next-generation deep sequencing are covered. Determining gene essentiality and statistical analysis on data collected are also discussed.