Figure 5

Functional assessment of barcoded primers by quantitative PCR and pyrosequencing analysis. A. Distribution of Ct scores for 96 barcoded LSU559 primers tested under identical reaction conditions. Bars represent relative frequency of occurrence of Ct scores. B. Q-Q plot of sample Ct scores versus those drawn from a theoretical normal distribution. Line denotes expected values for data obtained from a normal distribution. C. Scatterplot of Ct scores for each barcoded primer assayed in two separate PCR experiments and using different primer aliquots. The Pearson correlation coefficient of the two data sets is 0.48. B. Distribution of trimmed pyrosequencing reads for each of 47 barcoded LSU559 primers tested under indentical reaction conditions. Bars represent relative frequency of occurrence of sequence lengths (nts). E. Q-Q plot of pyrosequencing read lengths versus those drawn from a theoretical normal distribution. Line denotes expected values for data obtained from a normal distribution. F. Scatterplot of pyrosequencing read lengths vs. Ct scores for each barcoded primer assayed by pyrosequencing. The Pearson correlation coefficient of the two data sets is -0.01.