Week 15
From 2007.igem.org
(Difference between revisions)
| Line 33: | Line 33: | ||
::-pTet-LacI-T + I763007; | ::-pTet-LacI-T + I763007; | ||
::-pTet-LacI (low copy plasmid); | ::-pTet-LacI (low copy plasmid); | ||
| - | + | ::-I763032 (low copy plasmid); | |
| + | ::-I763007 (low copy plasmid). | ||
| + | *We inoculate in 6ml: | ||
| + | ::-I763031 (low copy plasmid); | ||
| + | ::-Plac-CI-Lacy (low copy plasmid). | ||
| + | *Ligation for: | ||
| + | ::-Plac-CI-Lacy (Eco/Spe)+GFP.(Eco/Xba). | ||
::'''10/10/07''' | ::'''10/10/07''' | ||
| - | + | *Miniprep of: | |
| - | + | ::-Plac-CI-Lacy (low copy plasmid); | |
| - | + | ::-I763031(low copy plasmid). | |
| - | + | *Digestion for : | |
| + | ::-I763031 with Xba/Pst1; | ||
| + | ::-Plac-CI-Lacy (low copy plasmid) with Spe/Pst1; | ||
| + | *Run on electrophoresis gel and Plac-CI-Lacy band extraction. | ||
| + | *Ligation for : | ||
| + | ::-Plac-CI-Lacy (low copy plasmid)+GFP; | ||
| + | ::-Ptet-LacI+pSB4A3; | ||
| + | ::-Ptet-LacI-LacI(high copy plasmid). | ||
| + | *We inoculate in 6ml: | ||
| + | ::-I763031 (low copy plasmid); | ||
| + | ::-Plac-CI-Lacy (low copy plasmid); | ||
| + | ::-pTet-LacI-T + I763007(high copy plasmid); | ||
| + | ::-I763007 (low copy plasmid); | ||
| + | ::-Ptet-LacI(low copy plasmid). | ||
::'''10/11/07''' | ::'''10/11/07''' | ||
| - | + | *Stock preparation (500ul) of: | |
| - | + | ::-I763031 (low copy plasmid); | |
| - | + | ::-Plac-CI-Lacy (low copy plasmid); | |
| - | + | ::-pTet-LacI-T + I763007(high copy plasmid); | |
| + | ::-I763007 (low copy plasmid); | ||
| + | ::-Ptet-LacI(low copy plasmid). | ||
| + | *Miniprep of: | ||
| + | ::-I763031 (low copy plasmid); | ||
| + | ::-Plac-CI-Lacy (low copy plasmid); | ||
| + | ::-pTet-LacI-T + I763007(high copy plasmid); | ||
| + | ::-I763007 (low copy plasmid); | ||
| + | ::-Ptet-LacI(low copy plasmid). | ||
| + | *Fluorescence test of: | ||
| + | ::-I763031 (low copy plasmid); | ||
| + | ::-pTet-LacI-T + I763007(high copy plasmid); | ||
| + | ::-I763007 (low copy plasmid). | ||
| + | *Control digestion of: | ||
| + | ::-I763031 with Xba/Pst1; | ||
| + | ::-Plac-CI-Lacy with Xba/Pst1 and Spe/Xba; | ||
| + | ::-pTet-LacI-T + I763007 with Xba/Pst1 and Spe/Xba; | ||
| + | ::-Ptet-LacI with Xba/Pst1; | ||
| + | ::-I763007 with Xba/Pst1. | ||
| + | *Run on electrophoresis gel and band extraction. | ||
| + | *Transformation of: | ||
| + | ::-Plac-CI-Lacy+GFP; | ||
| + | ::-Ptet-LacI-LacI; | ||
| + | ::-pLac-Lacy-GFP. | ||
| + | *We inoculate | ||
| + | ::-Plac-CI-Lacy+GFP. | ||
::'''10/12/07''' | ::'''10/12/07''' | ||
| - | + | *Miniprep of: | |
| + | ::-Plac-CI-Lacy+GFP. | ||
| + | *Digestion of: | ||
| + | ::-Plac-CI-Lacy+GFP with Xba/Pst1; | ||
| + | ::-Plac-CI-Lacy+GFP with Eco/Xba. | ||
| + | *Run on electrophoresis gel. | ||
[[Bologna | Back]] | [[Bologna | Back]] | ||
Revision as of 09:02, 12 October 2007
- 10/08/07
- We inoculate:
- -I763031 (low copy plasmid);
- -Plac-CI-Lacy (low copy plasmid);
- -E.Coli;
- -I763032 (high copy plasmid).
- Digestion of:
- -I763035 with Eco/Pst1;
- -I763007 with Eco/spe;
- -Ptet-LacI-T with Eco/Xba;
- -pSB4A3 with Spe/Xba.
- Ligation for:
- -I763035+pSB4A3;
- -I763007+Ptet-lacI-t;
- -I763007+pSB4A3;
- -I763032+pSB4A3.
- Run on electrophoresis gel.
- I763007 and Ptet-lacI-T band extraction.
- O/N ligation for:
- -Ptet-LacI-T+I763007;
- -Ptet-LacI+pSB4A3;
- -I763032+pSB4A3;
- -I763007+pSB4A3.
- We inoculate:
- -I763031 (low copy plasmid);
- -Plac-CI-Lacy (low copy plasmid);
- -I763032 (high copy plasmid).
- 10/09/07
- Transformation of:
- -pTet-LacI-T + I763007;
- -pTet-LacI (low copy plasmid);
- -I763032 (low copy plasmid);
- -I763007 (low copy plasmid).
- We inoculate in 6ml:
- -I763031 (low copy plasmid);
- -Plac-CI-Lacy (low copy plasmid).
- Ligation for:
- -Plac-CI-Lacy (Eco/Spe)+GFP.(Eco/Xba).
- 10/10/07
- Miniprep of:
- -Plac-CI-Lacy (low copy plasmid);
- -I763031(low copy plasmid).
- Digestion for :
- -I763031 with Xba/Pst1;
- -Plac-CI-Lacy (low copy plasmid) with Spe/Pst1;
- Run on electrophoresis gel and Plac-CI-Lacy band extraction.
- Ligation for :
- -Plac-CI-Lacy (low copy plasmid)+GFP;
- -Ptet-LacI+pSB4A3;
- -Ptet-LacI-LacI(high copy plasmid).
- We inoculate in 6ml:
- -I763031 (low copy plasmid);
- -Plac-CI-Lacy (low copy plasmid);
- -pTet-LacI-T + I763007(high copy plasmid);
- -I763007 (low copy plasmid);
- -Ptet-LacI(low copy plasmid).
- 10/11/07
- Stock preparation (500ul) of:
- -I763031 (low copy plasmid);
- -Plac-CI-Lacy (low copy plasmid);
- -pTet-LacI-T + I763007(high copy plasmid);
- -I763007 (low copy plasmid);
- -Ptet-LacI(low copy plasmid).
- Miniprep of:
- -I763031 (low copy plasmid);
- -Plac-CI-Lacy (low copy plasmid);
- -pTet-LacI-T + I763007(high copy plasmid);
- -I763007 (low copy plasmid);
- -Ptet-LacI(low copy plasmid).
- Fluorescence test of:
- -I763031 (low copy plasmid);
- -pTet-LacI-T + I763007(high copy plasmid);
- -I763007 (low copy plasmid).
- Control digestion of:
- -I763031 with Xba/Pst1;
- -Plac-CI-Lacy with Xba/Pst1 and Spe/Xba;
- -pTet-LacI-T + I763007 with Xba/Pst1 and Spe/Xba;
- -Ptet-LacI with Xba/Pst1;
- -I763007 with Xba/Pst1.
- Run on electrophoresis gel and band extraction.
- Transformation of:
- -Plac-CI-Lacy+GFP;
- -Ptet-LacI-LacI;
- -pLac-Lacy-GFP.
- We inoculate
- -Plac-CI-Lacy+GFP.
- 10/12/07
- Miniprep of:
- -Plac-CI-Lacy+GFP.
- Digestion of:
- -Plac-CI-Lacy+GFP with Xba/Pst1;
- -Plac-CI-Lacy+GFP with Eco/Xba.
- Run on electrophoresis gel.
