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Design
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Size of the transposon library
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- Genome size
- Resolution needed for experiments
- Polarity of transposon (bidirectional / unidirectional / neither)

Selection
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- Number of generations.

Replication
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- Repetition of the selective condition.

Bottlenecks in culture and during selection
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- Calculate and consider:
    - number of mutants in library
    - cell density of aliquots

*i.e., If your stored library is at a density of 5 x 10\ :sup:`7` CFU/ml, then taking 100 μl for an experiment is approximately 5 x 10\ :sup:`6` bacterial cells. If you original library contains 10,000 independent mutants this is likely ok. However, if your original library is 1,000,000 mutants then each aliquot will be subsampling the original library.*