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.
Solis BioDyne has always welcomed all innovative ideas with open hands. In order to elevate our mission and introduce new solutions to the field of genetic testing, we are delighted to present to you our OEM service offerings. Learn how you can implement the Stability TAG technology into your business.
In Estonia, we pretty much spend half a year in the dark and cold. It’s therefore very nice to have a person around who lights up the whole room and brings the warmth of summer with her everywhere she goes. Our own sunshine Narges was chosen by co-workers as the greatest Makes My Day Person in the company, so here is a little story about her to make your day.
Science is continually moving toward more accurate and individual-specific approaches when it comes to developing new genotyping technologies. In light of that, we would like to discuss high-throughput genotyping, which allows for a large number of precise results to be obtained fast.
Have you ever used chat on our website to talk about our products or PCR or whatever came to your mind? Have you ever asked for advice from our customer support? If you have, the chances are you were talking to Birgit. She is the little miracle behind your perfectly functioning experiment. Now you have the opportunity to get to know her a little better.