IGEM:MIT/2007/Notebook/2007-8-11

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(Agenda)
(Agenda)
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==Agenda==
==Agenda==
#Double digestion:
#Double digestion:
-
#*R0051.B0034, I14032.B0034, CPX.B0014 - for 3A assembly into pSB1AC3
+
#*R0051.B0034, I14032.B0034, CPX.B0014 - for 3A assembly into pSB1AC3.
#*F2620.B0034.CPX, B0014 - for standard assembly and to figure out extra bands in western blot
#*F2620.B0034.CPX, B0014 - for standard assembly and to figure out extra bands in western blot
#Purify digestion reactions. Remember to use <b>BUFFER TE</b> and NOT H2O.
#Purify digestion reactions. Remember to use <b>BUFFER TE</b> and NOT H2O.
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#*F2620.B0034.CPX.B0014
#*F2620.B0034.CPX.B0014
#Transform ligations into DH5a and plate.
#Transform ligations into DH5a and plate.
 +
 +
==Holding off on constitutive promoter digestion/purification/ligation/transformation until sequencing results come back==

Revision as of 12:25, 11 August 2007

Agenda

  1. Double digestion:
    • R0051.B0034, I14032.B0034, CPX.B0014 - for 3A assembly into pSB1AC3.
    • F2620.B0034.CPX, B0014 - for standard assembly and to figure out extra bands in western blot
  2. Purify digestion reactions. Remember to use BUFFER TE and NOT H2O.
  3. Ligations of digestion mixtures
    • R0051.B0034.CPX.B0014
    • I14032.B0034.CPX.B0014
    • F2620.B0034.CPX.B0014
  4. Transform ligations into DH5a and plate.

Holding off on constitutive promoter digestion/purification/ligation/transformation until sequencing results come back

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