design: For DNA templates, 15 ng of DNA is required per 100 base pairs in 12 ul volumes or less. DNA should be eluted in water for best results. For example, a 5 kb plasmid requires 750 ng of template DNA. For a 500 bp PCR product, 75 ng of template are required. Each primer constitutes one reaction, and primers should be provided at 10 uM concentration, 2 uL per reaction. Good quality starting template DNA should provide about 800-1000 bases of good quality sequence.