Bacterial growth calculator

Growth rate, doubling time and time to a target OD from two OD600 readings.

ReadingsLog phase, OD600 about 0.1–0.8

Result
growth rate μ (h⁻¹)
generations between readings

How to calculate bacterial growth rate from OD600

In log phase a culture grows exponentially, so the optical density at 600 nm rises as OD(t) = OD₀ · eμt. Two readings taken during that phase are enough to solve for thespecific growth rate μ:

μ = ln(OD₂ / OD₁) ÷ (t₂ − t₁)

μ has units of h⁻¹ (or min⁻¹). It depends on the strain, the medium, temperature and aeration, so measure it under the conditions you actually use rather than assuming a textbook value.

Doubling time and generation time

The doubling time, also called generation time, is how long the population takes to double. It follows directly from μ:

doubling time = ln(2) ÷ μ

The number of generations between the two readings is log₂(OD₂ / OD₁). For OD 0.1 → 0.4 that is 2 generations, so μ = ln(4) / 2 h = 0.693 h⁻¹ and the doubling time is 1.0 h. If you already know the doubling time and want to go the other way, use thedoubling time calculator.

When will the culture reach a target OD?

The most common use is timing the harvest: competent cells at OD 0.4–0.6, IPTG induction at OD 0.6–0.8, or a flask that must hit OD 1.0 before the next step. Extrapolating the same exponential curve gives

ttarget = t₂ + ln(ODtarget / OD₂) ÷ μ

The result is reported as elapsed time and as minutes after the second reading, so you can set a timer. The prediction assumes growth stays exponential; the closer the target is to the second reading, the more reliable it is.

Which OD600 readings to use

  • Take both readings in log phase, typically OD600 0.1 to 0.8. Readings during lag phase underestimate μ; readings near stationary phase overestimate the doubling time.
  • Keep readings above about 0.05 so instrument noise does not dominate the ratio.
  • Above roughly OD 1 most spectrophotometers are non-linear. Dilute the sample and multiply back, or the calculator will flag the reading.
  • Space the readings by at least one doubling. Two readings 10 minutes apart on a 40-minute doubler give a poor estimate.
  • OD is proportional to biomass, not cell number. To convert to cells/mL use theOD600 calculator.

Typical doubling times

Organism and conditionsDoubling time
E. coli, LB, 37 °C, shaking20–30 min
E. coli, M9 minimal glucose, 37 °C45–60 min
E. coli, LB, 30 °C40–60 min
Bacillus subtilis, LB, 37 °C25–40 min
Saccharomyces cerevisiae, YPD, 30 °C90–120 min
Mycobacterium tuberculosis18–24 h

Frequently asked questions

How do you calculate bacterial doubling time from OD600?

Take two OD600 readings in log phase. The specific growth rate is μ = ln(OD2 / OD1) / (t2 − t1), and the doubling time is ln(2) / μ. OD 0.1 to 0.4 over 2 hours gives μ = 0.693 h⁻¹ and a doubling time of 1.0 hour.

How do you predict when a culture reaches a target OD?

Assuming exponential growth continues, the time to the target is t = t2 + ln(OD_target / OD2) / μ, where μ is the growth rate from the two readings. For OD 0.4 at 2 h with μ = 0.693 h⁻¹, OD 0.6 is reached at 2.58 h, about 35 minutes after the second reading.

What OD600 range does the calculation apply to?

Only exponential (log-phase) growth, roughly OD600 0.1 to 0.8 for E. coli in rich medium. Below 0.05 readings are noisy, and above about 1 the spectrophotometer response is no longer linear and growth slows, so predictions drift.

What is a typical doubling time for E. coli?

About 20 minutes in LB at 37 °C with good aeration, 30 to 40 minutes in minimal medium, and longer at lower temperatures or in expression strains carrying a burdensome plasmid.

Is generation time the same as doubling time?

Yes. Generation time is the interval in which the population doubles. The number of generations between two readings is log2(OD2 / OD1).