A New Solution for Optimizing PCR Master Mix Liquid Transfers

A-New-Solution-for-Optimizing-PCR-Master-Mix-Liquid-Transfers

A-New-Solution-for-Optimizing-PCR-Master-Mix-Liquid-TransfersOverview

Measuring and ensuring accuracy and precision of liquid delivery steps when transitioning from handheld pipettes to automated liquid handlers is a constant challenge, especially for complex assays like qPCR and next generation sequencing. These assays often require several automation steps and utilize complex solutions (e.g. PCR master mix) that may pipette very differently than aqueous solutions. Determining an ideal liquid class for these complex solutions can be an arduous process made only more difficult by the fluid handling properties of the solutions themselves. In many cases, dialing in liquid class settings with the actual test solutions is problematic due to prohibitive cost, rarity, or other. For these types of high value test solutions, users often create an analog solution that hopefully has the same liquid handling characteristics as the sample it is supposed to mimic. A common example of such an analog is 20-50% glycerol/water as a mimic for PCR master mix. How well this analog works depends on how well it actually mimics the fluidic properties of the test solution.

Artel has developed PCRMix QualAssure, a test solution with liquid handling properties that are analogous to various commercially available PCR master mixes. PCRMix is a master-mix-like reference solution that can be used to assist in liquid class optimization, volume verification, and calibration of automated liquid handers (ALH). This solution is the latest in the Artel MVS product line and has been specifically engineered to reduce cost and time requirements in addition to generating the most accurate and reliable results for assays that contain master mix. In this study we show the similarities of PCRMix QualAssure compared to various commercially available PCR and qPCR master mixes. Equally, we show that aqueous and glycerol/water solutions are not suitable mimics for PCR master mix.