Run your PCR reaction with labeled primers. You may need to optimize your PCR conditions based on your primer concentration, annealing temperature, DNA template and product length. The quantity of the PCR products varies depending on the amount and quality of the DNA template used for the PCR reactions. In general, the signal of PCR product is too strong to be directly loaded on the ABI 3130. We recommend preparing a dilution series of the PCR products and running electrophoresis in order to optimize the fluorescence. The allele fluorescence intensities should fall between ~1000–4000 Relative Fluorescence Units (RFU). Peaks lower than ~300 RFU and higher than ~6000 RFU should be interpreted with caution.
1 uL of diluted sample (1:10, 1:20 1:30 and 1:40 ...... PCR product with water)
0.5 uL of LIZ size standard
15.5 uL of deionized formamide
Mix into each well of a 96-well plate compatible with the ABI 3130 instrument.