Mutagenesis of PQN-59, Removal of GFP

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#### 07/10/17 | 8:18 PM Performed mutagenesis reaction another time, since the first did not get the desired bands. 1. Added 5uL of template DNA 2. 1.25uL of each primer set, this time using `pqn-59_GFP_TOP` and `pqn-59_GFP_BOT` primer. 3. Added 1uL of HF polymerase 4. Added 5uL of 10X rxn buffer 5. Added 3uL of DMSO 6. Performed 11kb QuickChange cycle on PCR machine. 7. After this, added 1uL of DpnI to the mix and let it sit for one hour. 8. Ran this sample on gel, but did not get any distinct bands. #### 07/14/17 | 2:57 PM We have officially closed this cloning project, since we have repeatedly been unable to procure proper bands that show digestion. Instead we will use a preexisitng vecotr that featrues PQN-59 promoter, GFP and add a 3'UTR for proper processing.

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