IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/05/26
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Bootcamp Day 3
No growth observed on plates transformed with ligation products from yesterday. Re-plated 12M and 12O constructs as follows:
Incubate 10 minutes RT Inactivate 20 minutes 80°C
NB: used 5X Invitrogen ligase buffer with NEB ligase instead of 10X NEB ligase buffer which may be a problem...
100 ul competent cells, 20 ul ligation product
add 100 ul LB broth
37°C 1 hour recovery.
For both 12O and 12M:
100 ul on CAP
100 ul on LB
50 ul on CAP
50 ul on LB