Finding love for PCR

The running joke with PCR is that if something can go wrong, it will go wrong. Quite often it’s even impossible to determine why some samples turned out fine while the others did not. In a situation like this, it would be amazing to know some trick or a secret to avoid spending all the time and resources to do the experiment again. Here are a few we are willing to share so that you could find love for PCR.

Sometimes PCR goes wrong even before it starts. It’s important to have a good DNA sample, to be able to properly multiply it. If the DNA is not pure enough, the reaction might give you a messy answer. Contamination is therefore one of the biggest issues of PCR. Luckily, even if it turns out to be impossible to get pure DNA, certain methods can help prevent contaminants from ruining the experiment. One of these is using dUTP and UNG enzyme. UNG and dUTP enable the removal of residual products from previous PCR amplifications and so prevent cross-contamination.

When you start mixing all the necessary components together for PCR, you realize there are a lot of them and in different concentrations. In a situation like this, things can go wrong and it’s hard to say which component was at fault, what should be added more, what less, and what might have been contaminated. This kind of experimenting will certainly take up much of your time. The more things you have together in one master mix the smaller the chance of errors and contamination.

Once you are convinced that your DNA is pure, you don’t have contamination, all the concentrations are optimal, everything looks perfect and then you realize you forgot ice. Normal Taq DNA polymerase is active even at lower temperatures, therefore your reaction will start while you are still preparing. So, another trick to a successful PCR is choosing the right DNA polymerase. The preferred choice is hot-start DNA polymerase, which is activated only by pre-incubation at 95°C, preventing any unspecific polymerase activity at lower temperatures during reaction setup.

Another thing to be considered is time. It usually takes over an hour for the PCR reaction to finish, so a big part of your day will be spent on waiting while you could be already writing down the results. The truth is, with the right components, it can all be done a lot faster.

Now the simplest yet the best trick to a successful PCR is using Solis BioDyne’s products. All the previously mentioned problems can be easily avoided by using any of the products listed in our product lineup. Try them out and find the love for doing PCR experiments again.

If you would like some additional tips and tricks, check out our previous blog posts:

You may also always consult our troubleshooting guide.