Sample dilution factors

Do not try to run undiluted PCR products - most likely it leads to detector saturation and unusable data.

It is recommended to dilute single PCR products in ratios from 1:10 to 1:50 (sometimes up to 1:200) before running them. Prepare dilutions in Milli-Q water.

If you see clear bands on agarose gel, it is recommended to use dilution of 1:50 or more.
When pooling markers to form panels, first pool raw PCR products and dilute that cocktail to reach final dilution range of 1:10 to 1:50.

Keep the amount of size standard fixed from assay to assay - it is enough for succesful Fragment Analysis Run. Vary the amount or dilution of the PCR product in the sample if necessary.

ABI3730xl DNA Analyzer is based on electrokinetic injection. During the injection DNA fragments and all negative charged ions are transferred to the capillaries. Negative charged impurities which have light molecular weight will interfere with electrokinetic injection thus decreasing the signal intensities of the run.