Team:UCLA/Notebook/Honeybee Silk/13 July 2015

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Colony PCR of 7/12 Transformation

• The transformation was successful for the 3:1 ligated products plates. I picked 3 colonies off the plate and suspended each in 100uL of ddH2O.
• Using APEX Taq Red Mastermix

Component	Volume (out of 25L)
APEX Taq Red Mastermix	12.5uL
10mM primer 10	1.25uL
10mM primer 11	1.25uL
Transformed Cells	1uL
Nuclease Free Water	9uL

Testing competency of BL21 cells from 7/12

• Used puc 19 vector at 50 picograms/ul to test transformation efficiency.
• Added 2 ul of puc 19 to 50 ul of competent BL21 cells
• Used this NEB protocol to transform with our BL21 cells.
• Plated 10% of the cells on a warm amp plate and put plate in at 4 pm 7/13