These individual percentages were then used to calculate the overall proportion of each of the four fish species across all faecal samples. Due to the stochasticity Semaxanib associated with low copy number DNA and primer dimer accumulation above CT values of 34, all CT values recorded above this level were attributed a CT value of 34. This approach enables the target amplicon��s presence to be acknowledged, whilst still allowing for it to be expressed proportionally to the other fish species within that sample. To enable comparison of the qPCR and HTS datasets, the proportions of each of the four major fish species within each faecal GDC-0941 customer reviews sample as determined via HTS were considered to the exclusion of all other prey species detected. Using these data in conjunction with that obtained via qPCR, the Pearson productmoment correlation coefficient was calculated to determine the degree of correlation between the datasets. Additionally, individual paired sample t-tests for each major fish species were used to determine if there was a significant difference between the data obtained via both methods for any of the four major fish species. Samples that recorded CT values .34 were excluded from statistical analyses, due to the stochasticity of qPCR above this threshold. All statistical analyses were carried out using the program R. Using the cloning approach, a total of nine fish species were identified from 129 sequences, in 22 of the 47 samples collected during the Aug ��08-Sep ��09 sampling period. Samples deemed to have failed either yielded no amplifiable DNA, were severely compromised by inhibitors, or had target copy numbers that were considered too low to be reliable. The dominant prey species detected within these samples was H. vittatus, present in 32% of samples, followed by S. robustus, found in 20% of samples, with S. sagax, E. australis and Sardinella lemuru each found in 9.8% of samples . A number of other minor prey items were also identified, however they were found to represent a small proportion of sequences . Of the 52 samples collected during the Oct ��10-Dec ��10 sampling period, only 27 samples were deemed to have yielded DNA of sufficient quality free of inhibition that they could advance to HTS analysis. The two independent GS-Junior runs generated a total of 7810 DNA sequences.