1. 1 100 120 University of Oklahoma Genome Center4/14/12

2. 2 100 120 150 University of Oklahoma Genome Center4/14/12

3. 3 100 120 130 140 150 The line was extrapolated from 3 data points, 100, 120 and 150 ul of amplification primer University of Oklahoma Genome Center4/14/12

4. Suggested Optimal Amount of Amplification Primer (ul) for Various Average Sized Template DNAs (bp) 4 Average Template DNA Size (bp) Optimal Amount of Amplification Primer (ul) 300100 500120 700130 900140 1200150 University of Oklahoma Genome Center4/14/12

5. Most recent amplicon sequencing results 5University of Oklahoma Genome Center4/14/12 For this amplicon run: – we loaded 1800K beads made from a mixture of amplicons of ~300 and ~500 bases each (see slide 5) on each half of the plate (3600K beads total) using 100 ul of amplification primer on one half and 120 ul of amplification primer on the other half. This run yielded 1,232, 028 reads that passed the filter, with 445,121,246 total bases for an average read length of 362 bases/read. The average read length for each of the 4 sample sets is shown on the next slide.

6. Average Read length for ~500 and ~300 bp fragments 6 We conclude that both sets of sequence reads cover the full length of the respective amplicons University of Oklahoma Genome Center4/14/12

7. Read length distribution for 1 kb genomic DNA fragments 7University of Oklahoma Genome Center4/14/12