Is extension primer optimization really important for my iPLEX reactions?
• Yes! As oligo of higher molecular weights fly slower than the lower molecular weight oligos, you need to ensure that your extension primers of are as equal intensity as possible. The rule of thumb is that your lowest intensity peak is at least 50% of your highest intensity peak. You can choose to do a 2-step, 3-step, 4-step or linear regression to optimize your primers. The primer optimization method is located in Appendix A of the iPLEX Gold Application Guide.