Week 15
From 2007.igem.org
(Difference between revisions)
| Line 49: | Line 49: | ||
::-I763031 with Xba/Pst1; | ::-I763031 with Xba/Pst1; | ||
::-I763016 (low copy plasmid) with Spe/Pst1; | ::-I763016 (low copy plasmid) with Spe/Pst1; | ||
| - | *Run on electrophoresis gel and | + | *Run on electrophoresis gel and I763016 band extraction. |
*Ligation for : | *Ligation for : | ||
::-I763016 (low copy plasmid)+GFP; | ::-I763016 (low copy plasmid)+GFP; | ||
::-I763024+pSB4A3; | ::-I763024+pSB4A3; | ||
| - | ::-I763024 | + | ::-I763024+LacI(high copy plasmid). |
*We inoculate in 6ml: | *We inoculate in 6ml: | ||
::-I763031 (low copy plasmid); | ::-I763031 (low copy plasmid); | ||
| Line 59: | Line 59: | ||
::-I763026 + I763007(high copy plasmid); | ::-I763026 + I763007(high copy plasmid); | ||
::-I763007 (low copy plasmid); | ::-I763007 (low copy plasmid); | ||
| - | ::- | + | ::-I763024(low copy plasmid). |
::'''10/11/07''' | ::'''10/11/07''' | ||
*Stock preparation (500ul) of: | *Stock preparation (500ul) of: | ||
| Line 66: | Line 66: | ||
::-I763026 + I763007(high copy plasmid); | ::-I763026 + I763007(high copy plasmid); | ||
::-I763007 (low copy plasmid); | ::-I763007 (low copy plasmid); | ||
| - | ::- | + | ::-I763024(low copy plasmid). |
*Miniprep of: | *Miniprep of: | ||
::-I763031 (low copy plasmid); | ::-I763031 (low copy plasmid); | ||
| Line 85: | Line 85: | ||
*Run on electrophoresis gel and band extraction. | *Run on electrophoresis gel and band extraction. | ||
*Transformation of: | *Transformation of: | ||
| - | ::- | + | ::-I763019; |
| - | ::- | + | ::-I763024-LacI; |
::-pLac-Lacy-GFP. | ::-pLac-Lacy-GFP. | ||
*We inoculate | *We inoculate | ||
| - | ::- | + | ::-I763019. |
::'''10/12/07''' | ::'''10/12/07''' | ||
*Miniprep of: | *Miniprep of: | ||
| - | ::- | + | ::-I763019. |
*Digestion of: | *Digestion of: | ||
| - | ::- | + | ::-I763019 with Xba/Pst1; |
| - | ::- | + | ::-I763019 with Eco/Xba. |
*Run on electrophoresis gel. | *Run on electrophoresis gel. | ||
Revision as of 10:15, 12 October 2007
- 10/08/07
- We inoculate:
- -I763031 (low copy plasmid);
- -I763016 (low copy plasmid);
- -E.Coli;
- -I763032 (high copy plasmid).
- Digestion of:
- -I763035 with Eco/Pst1;
- -I763007 with Eco/spe;
- -I763026 with Eco/Xba;
- -pSB4A3 with Spe/Xba.
- Ligation for:
- -I763035+pSB4A3;
- -I763007+I763026;
- -I763007+pSB4A3;
- -I763032+pSB4A3.
- Run on electrophoresis gel.
- I763007 and Ptet-lacI-T band extraction.
- O/N ligation for:
- -I763026+I763007;
- -I763024+pSB4A3;
- -I763032+pSB4A3;
- -I763007+pSB4A3.
- We inoculate:
- -I763031 (low copy plasmid);
- -I763016 (low copy plasmid);
- -I763032 (high copy plasmid).
- 10/09/07
- Transformation of:
- -I763026 + I763007;
- -I763024 (low copy plasmid);
- -I763032 (low copy plasmid);
- -I763007 (low copy plasmid).
- We inoculate in 6ml:
- -I763031 (low copy plasmid);
- -I763016 (low copy plasmid).
- Ligation for:
- -I763016 (Eco/Spe)+GFP.(Eco/Xba).
- 10/10/07
- Miniprep of:
- -I763016 (low copy plasmid);
- -I763031(low copy plasmid).
- Digestion for :
- -I763031 with Xba/Pst1;
- -I763016 (low copy plasmid) with Spe/Pst1;
- Run on electrophoresis gel and I763016 band extraction.
- Ligation for :
- -I763016 (low copy plasmid)+GFP;
- -I763024+pSB4A3;
- -I763024+LacI(high copy plasmid).
- We inoculate in 6ml:
- -I763031 (low copy plasmid);
- -I763016 (low copy plasmid);
- -I763026 + I763007(high copy plasmid);
- -I763007 (low copy plasmid);
- -I763024(low copy plasmid).
- 10/11/07
- Stock preparation (500ul) of:
- -I763031 (low copy plasmid);
- -I763016 (low copy plasmid);
- -I763026 + I763007(high copy plasmid);
- -I763007 (low copy plasmid);
- -I763024(low copy plasmid).
- Miniprep of:
- -I763031 (low copy plasmid);
- -I763016 (low copy plasmid);
- -I763026 + I763007(high copy plasmid);
- -I763007 (low copy plasmid);
- -I763024(low copy plasmid).
- Fluorescence test of:
- -I763031 (low copy plasmid);
- -I763026 + I763007(high copy plasmid);
- -I763007 (low copy plasmid).
- Control digestion of:
- -I763031 with Xba/Pst1;
- -I763016 with Xba/Pst1 and Spe/Xba;
- -I763026 + I763007 with Xba/Pst1 and Spe/Xba;
- -I763024 with Xba/Pst1;
- -I763007 with Xba/Pst1.
- Run on electrophoresis gel and band extraction.
- Transformation of:
- -I763019;
- -I763024-LacI;
- -pLac-Lacy-GFP.
- We inoculate
- -I763019.
- 10/12/07
- Miniprep of:
- -I763019.
- Digestion of:
- -I763019 with Xba/Pst1;
- -I763019 with Eco/Xba.
- Run on electrophoresis gel.
