Imagine you are preparing your samples for PCR on a hot summer day. You have already mixed all the necessary components together, now you just have to insert your samples into the PCR machine. Suddenly your co-worker shows up and says that they need only a few minutes of your time to discuss a very important matter. As per usual a few minutes then turns into fifteen minutes. Finally, you turn to your samples and insert them into the PCR machine. A little while later it’s time to analyze the results, but for some reason, they are messy and don’t make sense. You sigh since you have wasted a perfectly good summer day, but gotten nothing out of it. So how to make sure this doesn’t happen again?
First, let’s talk about what went wrong. Presuming that there was no contamination and all the primers and PCR mix components were working properly, there was only the issue of letting your samples sit at room temperature for fifteen minutes. During that time your primers probably randomly bound to DNA templates or to each other forming primer dimers and starting non-specific amplification, because there was nothing to stop them from doing that. In the end, you ended up with lower yield and non-specific results, since part of the PCR efficiency went to amplifying unnecessary segments [1].
To avoid that the easy answer is using the hot start PCR technique. With this technique, you don’t have to worry about leaving your samples on the table while talking to a colleague or worry that once you get to the last samples the first ones might already be messed up. The main idea behind a hot start is that the reaction needs a hot start to work [1][2]. No non-specific amplification will occur during the reaction setup since the specially modified DNA polymerase won’t work until heated at 95 °C in the PCR machine [1][2]. So using the hot start PCR technique means that not only can you work calmly on your samples at a room temperature, but you will also end up with higher yields, better specificity, and sensitivity [1][2].
For your convenience, most Solis BioDyne PCR and qPCR mixes already contain reagents required for the hot start. We use chemical and oligo hot start methods to keep our enzymes inactive during reaction setup. In addition, due to Stability TAG technology, all our enzymes and master mixes have enhanced stability at room temperature with no activity loss for up to 1 month, so you don’t have to worry about not using ice while preparing your samples. Also, be sure to check out our SolisFAST® line products that are not only thermostable but so fast that you can repeat your perfect result and still have time left to enjoy the summer.
There are two options for target detection when doing qPCR - either you use dye or probe-based detection. Today we are going to take a deep dive into the world of dye-based qPCR, which was invented during the early nineties by Russell Higuchi [1] and is still used daily around the world.
Depression is a very complex disease, and it has many different aspects that affect it. Once we get to know more and more things about depression and its mechanisms, then in the end, we might discover new possible drugs or drug targets for treating depression.
If you ever need someone to help you with your computer, fix your house, join you in a sports competition, or just be a friend, Marek is your guy. Being one of the friendliest, most helpful people out there certainly makes him a really cool person everyone should know more about.
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.