Identifying essential genes in mycobacterium tuberculosis by global phenotypic profiling | Academic Article individual record
abstract

� Springer Science+Business Media New York 2015. 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.

author list (cited authors)
Long, J. E., Dejesus, M., Ward, D., Baker, R. E., Ioerger, T., & Sassetti, C. M.
publication date
2015
keywords
  • DNA Transposable Elements
  • Molecular Sequence Data
  • Bayes Theorem
  • Markov Chains
  • Gene Library
  • Chromosomes, Bacterial
  • Genes, Essential
  • Genomics
  • Phenotype
  • Mutagenesis, Insertional
  • Base Sequence
  • Mycobacterium Tuberculosis
  • Models, Statistical
  • DNA, Bacterial
altmetric score

3.5

citation count

30