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TUDelft/22 August 2008
From 2008.igem.org
22th August
DNA extraction test
Today another gel was run with DNA soaked from the spots. To test the soaking-protocol, various steps of the protocol were performed on the DNA. This time 10 ul of TE was used due to the soaking of the water in the punch, reducing free liquid greatly. Again various conditions were tested, and also the later sent spots from the promoter strength testing DNA were analyzed. To give an idea of DNA concentrations, all spots were nanodropped. These results are in the table below. For this testing we used a spot we won't be using and showed a good band on the gel of the repository. 1002 9A, a lock from Berkeley06, which had inconsistent sequence.
| Sample | ng/ul | A260/A280 |
|---|---|---|
| TE (also blanc) | 0.53 | 1.42 |
| 10 ul TE + 1002 9A punch, Tr | 4.94 | 0.69 |
| 10 ul TE + 1002 9A punch, Tr + spinned | 9.69 | 0.89 |
| 10 ul TE + 1002 9A punch, T = 42 | 11.04 | 0.75 |
| 10 ul TE + 1002 9A punch, T = 42 + Spinned | 11.78 | 1.23 |
| 10 ul TE + promoter strength punch, Tr | 5.94 | 0.97 |
| 10 ul TE + promoter strength punch, 42 C + spinned | 4.81 | 1.36 |
| 10 ul TE + 3 ul pSB1A7 (isolated on 20-08-08) | 11.69 | 1.66 |
These results have again rather low 260/280, but DNA concentrations that should be visible on a gel. Concentrations this time with increased following of iGEM procedure.
The gel showed no visible DNA bands for the soaked punches. The positive control did show bands, as can be seen below.
