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	*We inoculate:		*We inoculate:
	::-I763031 (low copy plasmid);		::-I763031 (low copy plasmid);
-	::-Plac-CI-Lacy (low copy plasmid);	+	::-I763016 (low copy plasmid);
	::-E.Coli;		::-E.Coli;
	::-I763032 (high copy plasmid).		::-I763032 (high copy plasmid).
Line 8:		Line 8:
	::-I763035 with Eco/Pst1;		::-I763035 with Eco/Pst1;
	::-I763007 with Eco/spe;		::-I763007 with Eco/spe;
-	::-Ptet-LacI-T with Eco/Xba;	+	::-I763026 with Eco/Xba;
	::-pSB4A3 with Spe/Xba.		::-pSB4A3 with Spe/Xba.
	*Ligation for:		*Ligation for:
	::-I763035+pSB4A3;		::-I763035+pSB4A3;
-	::-I763007+Ptet-lacI-t;	+	::-I763007+I763026;
	::-I763007+pSB4A3;		::-I763007+pSB4A3;
	::-I763032+pSB4A3.		::-I763032+pSB4A3.
Line 18:		Line 18:
	*I763007 and Ptet-lacI-T band extraction.		*I763007 and Ptet-lacI-T band extraction.
	*O/N ligation for:		*O/N ligation for:
-	::-Ptet-LacI-T+I763007;	+	::-I763026+I763007;
-	::-Ptet-LacI+pSB4A3;	+	::-I763024+pSB4A3;
	::-I763032+pSB4A3;		::-I763032+pSB4A3;
	::-I763007+pSB4A3.		::-I763007+pSB4A3.
	*We inoculate:		*We inoculate:
	::-I763031 (low copy plasmid);		::-I763031 (low copy plasmid);
-	::-Plac-CI-Lacy (low copy plasmid);	+	::-I763016 (low copy plasmid);
	::-I763032 (high copy plasmid).		::-I763032 (high copy plasmid).
Line 31:		Line 31:
	::'''10/09/07'''		::'''10/09/07'''
	*Transformation of:		*Transformation of:
-	::-pTet-LacI-T + I763007;	+	::-I763026 + I763007;
-	::-pTet-LacI (low copy plasmid);	+	::-I763024 (low copy plasmid);
	::-I763032 (low copy plasmid);		::-I763032 (low copy plasmid);
	::-I763007 (low copy plasmid).		::-I763007 (low copy plasmid).
	*We inoculate in 6ml:		*We inoculate in 6ml:
	::-I763031 (low copy plasmid);		::-I763031 (low copy plasmid);
-	::-Plac-CI-Lacy (low copy plasmid).	+	::-I763016 (low copy plasmid).
	*Ligation for:		*Ligation for:
-	::-Plac-CI-Lacy (Eco/Spe)+GFP.(Eco/Xba).	+	::-I763016 (Eco/Spe)+GFP.(Eco/Xba).
	::'''10/10/07'''		::'''10/10/07'''
	*Miniprep of:		*Miniprep of:
-	::-Plac-CI-Lacy (low copy plasmid);	+	::-I763016 (low copy plasmid);
	::-I763031(low copy plasmid).		::-I763031(low copy plasmid).
	*Digestion for :		*Digestion for :
	::-I763031 with Xba/Pst1;		::-I763031 with Xba/Pst1;
-	::-Plac-CI-Lacy (low copy plasmid) with Spe/Pst1;	+	::-I763016 (low copy plasmid) with Spe/Pst1;
	*Run on electrophoresis gel and Plac-CI-Lacy band extraction.		*Run on electrophoresis gel and Plac-CI-Lacy band extraction.
	*Ligation for :		*Ligation for :
-	::-Plac-CI-Lacy (low copy plasmid)+GFP;	+	::-I763016 (low copy plasmid)+GFP;
-	::-Ptet-LacI+pSB4A3;	+	::-I763024+pSB4A3;
-	::-Ptet-LacI-LacI(high copy plasmid).	+	::-I763024-LacI(high copy plasmid).
	*We inoculate in 6ml:		*We inoculate in 6ml:
	::-I763031 (low copy plasmid);		::-I763031 (low copy plasmid);
-	::-Plac-CI-Lacy (low copy plasmid);	+	::-I763016 (low copy plasmid);
-	::-pTet-LacI-T + I763007(high copy plasmid);	+	::-I763026 + I763007(high copy plasmid);
	::-I763007 (low copy plasmid);		::-I763007 (low copy plasmid);
	::-Ptet-LacI(low copy plasmid).		::-Ptet-LacI(low copy plasmid).
Line 63:		Line 63:
	*Stock preparation (500ul) of:		*Stock preparation (500ul) of:
	::-I763031 (low copy plasmid);		::-I763031 (low copy plasmid);
-	::-Plac-CI-Lacy (low copy plasmid);	+	::-I763016 (low copy plasmid);
-	::-pTet-LacI-T + I763007(high copy plasmid);	+	::-I763026 + I763007(high copy plasmid);
	::-I763007 (low copy plasmid);		::-I763007 (low copy plasmid);
	::-Ptet-LacI(low copy plasmid).		::-Ptet-LacI(low copy plasmid).
	*Miniprep of:		*Miniprep of:
	::-I763031 (low copy plasmid);		::-I763031 (low copy plasmid);
-	::-Plac-CI-Lacy (low copy plasmid);	+	::-I763016 (low copy plasmid);
-	::-pTet-LacI-T + I763007(high copy plasmid);	+	::-I763026 + I763007(high copy plasmid);
	::-I763007 (low copy plasmid);		::-I763007 (low copy plasmid);
-	::-Ptet-LacI(low copy plasmid).	+	::-I763024(low copy plasmid).
	*Fluorescence test of:		*Fluorescence test of:
	::-I763031 (low copy plasmid);		::-I763031 (low copy plasmid);
-	::-pTet-LacI-T + I763007(high copy plasmid);	+	::-I763026 + I763007(high copy plasmid);
	::-I763007 (low copy plasmid).		::-I763007 (low copy plasmid).
	*Control digestion of:		*Control digestion of:
	::-I763031 with Xba/Pst1;		::-I763031 with Xba/Pst1;
-	::-Plac-CI-Lacy with Xba/Pst1 and Spe/Xba;	+	::-I763016 with Xba/Pst1 and Spe/Xba;
-	::-pTet-LacI-T + I763007 with Xba/Pst1 and Spe/Xba;	+	::-I763026 + I763007 with Xba/Pst1 and Spe/Xba;
-	::-Ptet-LacI with Xba/Pst1;	+	::-I763024 with Xba/Pst1;
	::-I763007 with Xba/Pst1.		::-I763007 with Xba/Pst1.
	*Run on electrophoresis gel and band extraction.		*Run on electrophoresis gel and band extraction.
	*Transformation of:		*Transformation of:
-	::-Plac-CI-Lacy+GFP;	+	::-I763016+GFP;
	::-Ptet-LacI-LacI;		::-Ptet-LacI-LacI;
	::-pLac-Lacy-GFP.		::-pLac-Lacy-GFP.

---

Revision as of 09:43, 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 Plac-CI-Lacy 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);

-Ptet-LacI(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);

-Ptet-LacI(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:

-I763016+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.

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