Phenotype Testing and SDS- PAGE Mod2 Day4 Thursday Oct 16th, 2008.

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Phenotype Testing and SDS- PAGE Mod2 Day4 Thursday Oct 16th, 2008

First let’s look at colony PCR results

SDS-PAGE You will make a whole cell extract from equal # of cells --> OD600 Glass beads/SDS/BME/boiling steps Gel separates proteins based on size May want to change lane order for + control After 1 hour run, blot to nitrocellulose and block with milk solution until next week.

Yeast Phenotype Testing Choose plates you will use and put them in the incubator to dry out before running your protein gel Spot test demo…

For next time… What size is the protein you’re looking for (TAP = +20KDa) Make a figure of colony PCR result Calculate number of cells for RNA extraction