Project:Plant species testing

We are using primers for plants to test and refine our DNA extraction / PCR technique. First project is to test for peas. Inspiration in this paper.

First extraction and PCR - 17/7/12

We used two methods of extraction. Method 1:

• Blend peas with salt and detergent
• Incubate for 15 mins at 60 C (optional step not taken)
• Filter and centrifuge - recover supernatant
• Add proteinease K (0.8 ul - incubate for 1 hr at 50C then 94 C for 10 mins (inactivation of proteinase K)
• Precipitate with rubbing alcohol and meat tenderiser as an enzyme
• Centrifuge and remove supernatant to leave pellet

Method 2:

- 'Homogenize' with a pestle in 5% chelex solution for 1 min - Vortex for 10s then boil for 5 mins, then vortex again for 10s - Centrifuge for 1 min - recover supernatant and use as template

Using the PCR machine:

For soak: (

For PCR:

• File 4 enter
• Step
• temp
• time
• temp a
• time
• time e
• Segment 4 - 0 C for 0 seconds
• Cycle count
• Link to stored file - 20 C (to keep