Tips for PCR

Everyone prepares samples for PCR a little differently, but here are some tips and tricks that will help you get the best results.
 * 1) Make a master mix! (If you're not already using a pre-made one)
 * 2) Mix water, buffer, dNTPs, and polymerase in a tube that you can aliquot into your sample tubes. This can reduce the amount of pipetting errors made during the experiment.
 * 3) Add the mix to your control tubes first, then add the DNA for your samples. This can get a little bit complicated if you are using multiple types of DNA, so it can be easier sometimes to separate your mix.
 * 4) Once you add DNA, you can add your mix to your remaining tubes. All you have left are primers!
 * 5) Bubbles are bad. Try to avoid making any while you are preparing the tubes. Centrifuging your samples can get rid of any you may have made. Try mixing by tapping the tubes with your fingers.
 * 6) When pipetting a new liquid into a mixture, rinse the pipet tip by pipetting up and down gently a couple times. Try not to make bubbles! This will make sure you get all of the liquid out of your pipet. This can make a big difference if you are pipetting small amounts.
 * 7) Always add the largest amount of liquid first so you have a larger volume to work with. Usually in PCR, adding water should come first. This allows you to do the previous tip without fear of losing sample.
 * 8) Keep your samples on ice as soon as the DNA and polymerase are in the same tube. This will keep nuclease activity low.
 * 9) Add your samples to the thermocycler when the lid and sample wells have been preheated, if you are using a hot-start polymerase. This prevents potential exonuclease activity that could cut up your sample.