Biomod/2011/LMU/FolD'N'Assemble/Protocols: Difference between revisions
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Then we used electrophoresis in | Then we used electrophoresis in 2% agarose gel to test the samples | ||
====05-16-11 (Timon, Alex, Ralf)==== | ====05-16-11 (Timon, Alex, Ralf)==== | ||
Another test for best MgCl concentration and annealing time. We used a | Another test for best MgCl concentration and annealing time. We used a 2% agarose gel. | ||
[[Image:20110516.JPG|thumb|05-16-11, From left to right: ladder, scaffold, sample 1-6]] | [[Image:20110516.JPG|thumb|05-16-11, From left to right: ladder, scaffold, sample 1-6]] | ||
{| {{table}} | {| {{table}} | ||
Revision as of 08:03, 24 May 2011
May
05-13-11(Timon, Alex, Ralf)
We wanted to find the best MgCl concentration and the best annealing time for our construct.
| SampleNr. | Annealing time | MgCl(mM) | |
|---|---|---|---|
| 1 | 4h | 10 | |
| 2 | 4h | 14 | |
| 3 | 4h | 18 | |
| 4 | 24h | 10 | |
| 5 | 24h | 14 | |
| 6 | 24h | 18 | |
| 7 | 24h | 14 | dimers(2b) |
| 8 | 55h | 10 | |
| 9 | 55h | 14 | |
| 10 | 55h | 18 |
Then we used electrophoresis in 2% agarose gel to test the samples
05-16-11 (Timon, Alex, Ralf)
Another test for best MgCl concentration and annealing time. We used a 2% agarose gel.
| SampleNr. | Annealing time | MgCl(mM) | |
|---|---|---|---|
| 1 | 55h | 16 | |
| 2 | 55h | 18 | |
| 3 | 55h | 16 | dimers(2b) |
| 4 | 55h | 18 | dimers(2b) |
| 5 | 55h | 16 | dimers(4b) |
| 6 | 55h | 18 | dimers(4b) |
05-17-11(Timon, Alex, Ralf)
We took first TEM (transmission electron microscope) pictures of our construct.
05-18-11(Timon, Alex, Ralf)
New samples for next week: 200μL, 18mM MgCl, 55h
05-23-11(Timon, Alex, Ralf)
We prepared buffers with different pH.
- pH 4,5
- pH 5,03
- pH 5,8
- pH 7,8
05-24-11(Alex, Ralf, Timon)
- prepared 3 samples of origamis for measuring its stability in a changing pH environment
- exchanged standard buffer with pH adjusted buffer using centrifugal filters
- 3 samples, ~24 h exposure to pH values of 4.5, 5 and 6, 1x TE buffer, 18mM MgCl2
