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 Plac-CI-Lacy band extraction.
+
*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-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);
Line 59: Line 59:
::-I763026 + 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).
::'''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);
-
::-Ptet-LacI(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:
-
::-I763016+GFP;
+
::-I763019;
-
::-Ptet-LacI-LacI;
+
::-I763024-LacI;
::-pLac-Lacy-GFP.
::-pLac-Lacy-GFP.
*We inoculate  
*We inoculate  
-
::-Plac-CI-Lacy+GFP.     
+
::-I763019.     
::'''10/12/07'''
::'''10/12/07'''
*Miniprep of:
*Miniprep of:
-
::-Plac-CI-Lacy+GFP.
+
::-I763019.
*Digestion of:
*Digestion of:
-
::-Plac-CI-Lacy+GFP with Xba/Pst1;
+
::-I763019 with Xba/Pst1;
-
::-Plac-CI-Lacy+GFP with Eco/Xba.
+
::-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.


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