To help you get into the multiplexing workflow efficiently, there are a few key points to consider:
Figure 1. SolisFAST® Probe qPCR Mix (ROX) was used in a 4-plex probe-based qPCR with fourteen fold serial dilutions of human gDNA (40 ng –40 pg per reaction, three replicates at each concentration). qPCR was performed on a QuantStudio™ 6 Flex qPCR cycler (Applied BioSystems™) using ROX dye for normalization.
Using regular qPCR mixes in highly multiplexed assays carries a number of risks, such as late Ct values, low efficiency, low sensitivity, and low fluorescence. As regular qPCR mixes do not have a sufficient amount of components for amplifying more than 2 targets simultaneously, you are running the risk of getting false-negative results. Using specifically designed multiplex mixes will greatly increase your success.
For fast cycling speed and up to 5-plex reactions:
For standard cycling speed and up to 4-plex reactions:
By using the right product designed for multiplexing, you can rest assured you will enjoy similar sensitivity to singleplexing with a greatly reduced workload.
Figure 2. HOT FIREPol® Multiplex qPCR Mix (Purple) was used to perform 4-plex or single-plex probe-based qPCR with five tenfold serial dilutions of human cDNA (cDNA concentration ranges from 200 ng to 20 pg per reaction). Reactions were performed with Applied BioSystems™ QuantStudio™ 6 cycler using Purple dye for normalization.
This year seemed to just fly by because there was just so much happening all the time. It doesn’t feel that long ago when we did the recap for the year's first half. Now it’s time to focus on the second half of the year and all the wonder and joy we got to experience. As it is wintertime, feel free to grab a warm beverage and some gingerbread while you read.
In November of this year, the world population reached an astounding 8 billion people. To support the accompanying increase in food demand, beef and sheep farmers are using scientific interventions for assistance.
Contamination can ruin your day or even week. Data is not valid, a lot of cleaning must be done and supplies are wasted. Precious time and even samples can be lost. Prevention is the best strategy!
To give you a better insight into our SolisFAST® qPCR range’s inhibitor tolerance capabilities, we have put together a white paper. It includes results from different tests, shows the real-life application possibilities and feedback from our partners about the SolisFAST® qPCR products.