Expected value of the estimates obtained from pooled samples depending on the pool size n: Watterson's θ and Tajima's π. True value θ = 10 (green line). There is a considerable bias, if n is small compared to kλ, illustrating the need to use a bias correction with the estimates. Solid black line, λ = 30; red dashed line, λ = 5. (For Tajima's π, the bias does not depend on λ.)