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Team:UNIPV-Pavia/Notebook/Week19
From 2008.igem.org
(Difference between revisions)
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<br> | <br> | ||
*Sequencing results: | *Sequencing results: | ||
| - | *Lig.30 - we had another mutation in luxR coding sequence...no comments... | + | *Lig.30 - we had another mutation in luxR coding sequence...no comments...(position 349 of C0062, G->C, aminoacid changes...) |
| - | *Lig.34 - | + | *Lig.34-1 - OK |
| - | *We decided to test qualitatively | + | *Lig.34-3 - OK |
| - | ** | + | *Lig.34-4 - OK |
| - | ** | + | *Lig.34dil-1 - OK |
| + | *Lig.34dil-2 - OK | ||
| + | *Lig.35-3 - OK | ||
| + | *Lig.35-4 - OK | ||
| + | *Lig.35dil-1 - OK | ||
| + | *Lig.35dil-4 - OK | ||
| + | |||
| + | *Comments: Lig.34 and Lig.35 sequences were correct, but we have to notice that they are long parts, so their sequencing could not be extended to all the nucleotides. In particular, the mutated nucleotide of Lig.30 was not included in Lig.34 and Lig.35 sequencing. For this reason, we expect to find in Lig.34 the same G->C mutation as Lig.30. | ||
| + | |||
| + | *We decided to test qualitatively and quantitatively the mutated construct: | ||
| + | **T5 (R0051-B0030-C0062-B1006-R0062-B0030-E0040-B1006, that carries the G->C mutation) | ||
| + | *So, we infected 9 ml of LB + Amp with 30 ul of T5 glycerol stock. We incubated the culture at 37°C, 220 rpm overnight. | ||
| + | |||
| + | '''09/23/08''' | ||
| + | <br> | ||
| + | *We performed the experiment described in TEST 9 and quantitativeTEST 1 (The Project section, Experiments). | ||
| + | |||
| + | *We infected 9 ml of LB + Amp with 30 ul of Lig.31 and Lig.36 glycerol stocks. We incubated the cultures at 37°C, 220 rpm overnight. | ||
| + | |||
| + | '''09/24/08''' | ||
| + | <br> | ||
| + | *Data processing for 09/23/08 experiment. | ||
| + | |||
| + | *Glycerol stocks/miniprep for Lig.31 and Lig.36. | ||
| + | |||
| + | *We sent purified plasmids to Primm for sequencing. | ||
Revision as of 20:00, 26 October 2008
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Notebook
| Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 |
|---|---|---|---|---|---|---|
| Week 8 | Week 9 | Week 10 | Week 11 | Week 12 | Week 13 | Week 14 |
| Week 15 | Week 16 | Week 17 | Week 18 | Week 19 | Week 20 | Week 21 |
Week 19: 09/22/08 - 09/26/08
09/22/08
- Sequencing results:
- Lig.30 - we had another mutation in luxR coding sequence...no comments...(position 349 of C0062, G->C, aminoacid changes...)
- Lig.34-1 - OK
- Lig.34-3 - OK
- Lig.34-4 - OK
- Lig.34dil-1 - OK
- Lig.34dil-2 - OK
- Lig.35-3 - OK
- Lig.35-4 - OK
- Lig.35dil-1 - OK
- Lig.35dil-4 - OK
- Comments: Lig.34 and Lig.35 sequences were correct, but we have to notice that they are long parts, so their sequencing could not be extended to all the nucleotides. In particular, the mutated nucleotide of Lig.30 was not included in Lig.34 and Lig.35 sequencing. For this reason, we expect to find in Lig.34 the same G->C mutation as Lig.30.
- We decided to test qualitatively and quantitatively the mutated construct:
- T5 (R0051-B0030-C0062-B1006-R0062-B0030-E0040-B1006, that carries the G->C mutation)
- So, we infected 9 ml of LB + Amp with 30 ul of T5 glycerol stock. We incubated the culture at 37°C, 220 rpm overnight.
09/23/08
- We performed the experiment described in TEST 9 and quantitativeTEST 1 (The Project section, Experiments).
- We infected 9 ml of LB + Amp with 30 ul of Lig.31 and Lig.36 glycerol stocks. We incubated the cultures at 37°C, 220 rpm overnight.
09/24/08
- Data processing for 09/23/08 experiment.
- Glycerol stocks/miniprep for Lig.31 and Lig.36.
- We sent purified plasmids to Primm for sequencing.
