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| ### Using spike-in counts | |
| But we don't actually know the true fold-expansion of the reference strain, since | |
| it's not directly observed. However, a non-growing fitness-zero control can help, | |
| such as a heat-killed strain or a spike-in plasmid. | |
| We start with the equation before, | |
| $$ | |
| \log \left( \frac{c_i(t)}{c_{wt}(t)}\frac{c_{wt}(0)}{c_i(0)} \right) = \left(\frac{w_i}{w_{wt}} - 1 \right) \log \frac{n_{wt}(t)}{n_{wt}(0)} | |
| $$ | |
| But for the fitness-zero control, $w_{spike} = 0$, so: | |
| $$ | |
| \log \left( \frac{c_{spike}(t)}{c_{wt}(t)}\frac{c_{wt}(0)}{c_{spike}(0)} \right) = -\log \frac{n_{wt}(t)}{n_{wt}(0)} | |
| $$ | |
| This means that, although we don't know how the reference strain grows directly, its | |
| growth is given from the ratio of the spike counts to the reference counts, normalized | |
| to the same ratio at time 0. | |
| This leaves us with the overall equation: | |
| $$ | |
| \log \left( \frac{c_i(t)}{c_{wt}(t)}\frac{c_{wt}(0)}{c_i(0)} \right) = \left(1 - \frac{w_i}{w_{wt}} \right) \log \left( \frac{c_{spike}(t)}{c_{wt}(t)}\frac{c_{wt}(0)}{c_{spike}(0)} \right) | |
| $$ | |
| If we plot the left hand side against the right, we should get a straight line for | |
| each strain with intercept zero and gradient $1 - \frac{w_i}{w_{wt}}$. | |