1. Estimation of national population size can be important for setting conservation priorities but its methodology has received little critical attention. Sites for highly aggregated species are often prioritised if they contain 1% of national or biogeographical populations but the utility of this approach for other species is unclear. 2. To make recommendations for study design, we present methods used to estimate the UK population size of the common nightingale Luscinia megarhynchos. We assess the sensitivity of the population estimate to the analytical method used and identify sites of national importance for this territorial songbird. 3. Survey effort was directed by prior knowledge of the species’ distribution and the survey design maximised detectability by focussing on the period of greatest song output. We used three different statistical methods to account for detectability, estimating that 55–65% of the national population was detected during surveys. 4. Birds in areas not known to contain the species accounted for 13–23% of the population estimate. Methods to account for these individuals contributed the greatest uncertainty to the results, due to the difficulty of surveying a very large sample of random sites and consequent need to stratify the sample. 5. The 12 derived estimates ranged between 5094 and 5938 territorial males, with the confidence limits ranging from 4764 to 6534. Site delimitation, using clustering based on nearest-neighbour distances, identified one site clearly of national importance and several others potentially nationally important, depending on the population threshold and clustering distance used. 6. Synthesis and applications. National population estimation is difficult and requires that species-specific variability in detectability and individuals present outside surveyed areas are accurately accounted for through survey design and statistical analysis. Accounting for these sources of error will not always be possible and will hamper efforts to assess true population size and consequently to determine whether sites, however defined, exceed critical thresholds of importance. Resources may be better invested in other activities, for example in generating population trends based on relative indices. The latter are generally easier to produce, potentially more robust and arguably more suitable for many conservation applications.