Success! PCR worked!
- When we finished yesterday we had just loaded the samples into the PCR machine- basically we are trying to amplify or to produce many copies of our DNA sample. Why? What is the purpose? In this instance- you have a number of very similar looking bats and you want to identify them as being of the same species or not. Specific DNA samples are isolated and purified, PCR is used to amplify the strands. Finally, load the DNA into a gel with a dye and connect the well to the electricity. The DNA will move through the gel, the gel is then placed under fluorescent light and an image is captured. If the bats are of the same species they will have bands in all the same positions. If they are different species a band appearing in a position unique to that sample will be visualised. There you have it! This is part of what is being studied in the Bat lab. Species Identification.
- So this morning Clare and I made the gel for the electrophoresis.
- The samples were prepared: 2ul of dye and 8ul of sample. Samples were loaded into the gel wells, the electricity was switched on and the DNA moved from the negative end to the positive end of the gel. At the end of the process the gel was removed and placed into the chamber with the fluorescent light and the bands were captured on PC.
- This seems very easy but the preparation of the sample and making the PCR buffer is so important with a great number of individual steps, it is very time consuming. One small mistake which can be made without you noticing would not be picked up until the electrophoresis stage!
- This afternoon Clare and I are preparing samples for PCR from scratch, it’s all down to us!
- So we will know tomorrow if this afternoons work was up to scratch!
- Day 3, Clare and I loving the experience and our tutor Dr.Lao.
- Will post tomorrow and let you know how we did.