Plant DNA PCR

We set up a PCR reaction using the 5 plant DNAs that we extracted last week. We used two markers for the PCR: the ITS and psbA.

We clearly labelled two strips of 5 PCR tubes (so that’s 10 total, 5 tubes per strip) with our sample ID and the marker. Pipette 1 microlitre of your plant DNA into the respective PCR tubes, close the lid and place it in eyes.

Next, we prepared two MasterMixes (one for ITS and one for psbA). The steps are as followed:

  1. Label two 1.5mL centrifuge tube, one as MM psbA, the other as MM ITS.
  2. Thaw the reagents
  3. Add 15 microlitres distilled water, 2 microliteres buffer, 1 microlitre BSA, 0.2 microlitres dNTPs and F-Primer and R-Primer, 0.04 microlitres Taw and 1 microlitres of ITS.
  4. Repeat step 3 but with psbA.

Now that the MasterMixes have been created, pipette 19 microlitres of the MasterMix into the respective samples (if the PCR tube is labelled ITS, pipette ITS. If not, pipette psbA). Close the lid tightly, mix the tube and place it in the PCR machine.

Leave a Reply

Your email address will not be published. Required fields are marked *

Viewing Message: 1 of 1.
Warning

Important: Read our blog and commenting guidelines before using the USF Blogs network.

Skip to toolbar