new plan for the plasmid

so there's only one restriction enzyme that cuts in the plasmid in the insert and not the vector and i need two enzymes that do that. so sad. therefore, i get to make my own restriction site in the insert so that another enzyme can cut there and the insert can be gapped. so much work for such a little thing.

anyway, i get to use site-directed mutagenesis to create point mutations that change the dna sequence to have the specific site needed for the enzyme to cut. this is what site-directed mutagenesis using pcr looks like:



isn't that fun. again i just get to do all the set up and then the pcr does all the work for me. then i take the hopefully mutagenized plasmid, get rid of all the unmutagenized plasmid by chewing it up with an exonuclease, and then transform the mutagenized plasmid into bacterial cells so they can grow and amplify the amount of plasmid. then i get to mini-prep, digest, and run a gel to make sure it all worked out alright. whew. i may actually be busy in a few days and that is exciting!

tonight is bruce almighty on flagstaff. the movie really doesn't matter as much as the company, but it's supposed to be a funny movie anyway. i am bringing lots of cookies with me and that is also good.

i'm also excited to hang out with people this weekend. yay! it'll be weird though with alex gone and gerrit away. fewer people around, a smaller group. eh, it'll still be fun!

i have to run to campus now and pick up my package and get toliet paper from housing. how exciting!