racking Buffer is optional. If you choose to add Tracking Buffer to the master mix, 50 uL can be added directly to 1 mL of 2X Forget-Me-Not Universal Probe Master Mix, or add 0.5 uL per 20 uL reaction when setting up the assay master mix. Or Tracking Buffer may be added to your template samples at a final reaction concentration of 1X. For example, dilute 40XTracking Buffer to 20X in PCR grade water. Make 1:1 dilution of template with the 20X Tracking Buffer and add 2 uL of the mixture to the reaction