Doxycycline was added during seeding at a concentration of 0.2 mg/ml. The fluorescence was read 72 h after seeding . The readings obtained from 72 h after seeding were normalized to the value obtained for the untreated control knockdown condition for that cell line. DNA-based dietary analysis of faecal material has emerged as a promising tool to study animal biology, ecology and archaeology . Dietary analysis is not limited to the discovery of what an animal consumes; it can also give an insight into ecosystem health , species�� responses to environmental/anthropogenic stresses , and assist in the development of targeted strategies for conservation . It is evident from the increase in the use of genetic techniques that there is a Wortmannin distributor growing appreciation of the use of DNA-based faecal methods to investigate diet. The analysis of faecal material has proven to be a welcome move away from more invasive techniques used to study animal diet such as lethal sampling and stomach flushing , both of which have undesirable effects on the sampled population . Moreover, a general move towards molecular based approaches, e.g. fatty acid, stable isotope or DNA analysis, has allowed a shift from more subjective morphological approaches . The extraction and sequencing of DNA from faecal samples is seen to be an effective and reliable indicator of species�� diet, offering increased specificity and taxonomic resolution compared to other techniques . The possibility of misidentification of species is greatly reduced and the ability to account for a wider range of species within the actual diet is greatly increased when compared to morphology which relies entirely on analysis of undigested remains, therefore neglecting prey that may leave little trace of its consumption . DNA based quantitative estimates of diet, however, are not without problems. Issues have arisen as a result of primer biases and the problem of differential digestion still remains. Put simply, ����is what goes in what comes out���� ? Moreover, variability in the amount of DNA per unit biomass between species and different tissues is also difficult to quantify. Attempts to address such concerns have recently become an active area of research. Such efforts include; the use of blocking primers to circumvent the issue of predator DNA amplification ; the use of captive feeding trials to examine differential digestion; and the introduction of correction Vorinostat factors to account for DNA amount variability within species and tissues .