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From 2011.igem.org

Thuesday

Colonies on A2 but not on A1. Sekvenses back on B1 - we have a mutated version! Hurra.

Lab Work

• We took the A2'sout and put it in and overnight culture
• We transformed XL1-Blue with the A1'a we mutated friday, but this time we used 40 ul instead of 10 ul.
• We purified A2 with Mini prep from 2 different colonies.
• We analyzed A2 cut with Pst1 (which cut at the restrictionsite we want to remove)on an agarose gel. The results were incunclusive regarding the mutation, but we keep our fingers crossed and have sent them both to be sequences.
• Digested the standard plasmid from iGEM pSB1C3 with EcoR1 and Pst1
• We purified the digested plasmid by two methods -

- half of the plasmid was purified with gel purification - the other half was submitted to heatshock at 80 degress in order to kill the restrictionenzymes.