Pseudomonas isolate B3
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.