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 the Nobel Prize in Physiology or Medicine was awarded to Victor Ambros and Gary Ruvkun, two scientists credited with discovering microRNA and its role in post-transcriptional gene regulation. Now, 30 years after their finding, you can do microRNA experiments with ease by using our products designed to make discovering new things simple and hassle-free.
This summer we got to collaborate with a fun project organized by the MINT Campus in Germany. Not only does MINT campus inspire children and young people about these topics but it also introduces young people to sustainable, innovative developments in current research and technology.
Whether you are studying the genetic material of plants, brains or viruses, the experiment usually starts with extracting RNA from the sample material. It would be incredibly useful to get all the RNA extracted instead of it getting destroyed by the RNases before even starting the cDNA synthesis step. But how can we protect the RNA when RNases are all around us? Let’s find out!
There aren’t many people who can lead teams as well as our Head of Quality and Product Management Eva-Maria does. She is like that red string on a mystery board connecting all the departments. If you would like to get a little closer to solving the great leadership puzzle that Eva-Maria has mastered, have a look at the following story.
