Application Note: How to Pipette PCR Master Mix for Increased Accuracy in qPCR Results

Commonly Asked Questions  

• How can pipetting affect qPCR results?  - Because PCR Master Mix contains proteins, improper pipetting can lead to the creation of bubbles in the sample, which can skew the results of the experiment. 
• Which pipetting technique is best when using PCR Master Mix? - According to ISO guidelines, it is recommended to pre-wet the pipette tip 3 to 5 times, it is also recommended to use reverse pipetting technique as PCR Master Mixes are viscous. 
• Is pipette calibration important when using PCR Master Mix? - There is a propensity for error when dealing with viscous materials such as PCR Master Mix. Therefore, it is important to eliminate as many risk factors as possible. Ensuring your pipette is regularly maintained and calibrated is an effective way to keep results accurate and reproducible.
• Should I use a multichannel pipette for preparation of qPCR assays? - Using a multichannel pipette for qPCR assays ensures equal amounts of PCR Master Mix will be evenly and quickly distributed into several wells at once. This is a time-saving factor especially when working with a 96 or 384 well plate. Reproducibility can be increased in experiments further by using an electronic pipette and multi-dispensing mode.