Difference between revisions of "Project:Plant species testing"
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== First extraction and PCR - 17/7/12 == | == 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 | ||
| + | - 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 | ||
Revision as of 19:52, 17 July 2012
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 - 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