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Lab July 23 2009
From 2009.igem.org
No growth observed on our LB+amp plate from last night. Either the pUC18 plasmid was not done correctly or the cells were not properly competent. Greatest likelihood is that the cells were not competent.
Rather than try to debug our openwetware protocol, we've decided to revert back to the Preparation of Competent Cells Protocol used in BCMB 301 that the TA's are familiar with.
In preparation for that, we created 500ml of LB broth and sent it to be autoclaved.
Next, we are preparing the following stock solutions in 100 mL quantities:
1M NaOAc, pH 5.6 = 8.2g in 100 mL
1M MnCl2 = 19.79g in 100 mL
.25M NaCl = 1.46g in 100 mL
1M CaCl2 = 14.7g in 100 mL
These were sent for autoclaving to be used in the morning.
in 100ml of sterile LB, we placed one colony from our DH5α plate and placed it in a 250 ml erlenmeyer at 37 degrees under medium agitation.
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