Today transformed cells to introduce plasmids inside. In this case the cells were transformed Ura - and thus can not grow in media without uracil . He plasmid introduce them given the ura gene 3 with the ability to generate their own uracil therefore join those who will survive in the uracil -.
To accomplish all this:
-strains grew on rich medium plates
- cells resuspended in 1 ml of sterile water and centrifuged
-discard the supernatant and resuspended in the liquid remaining
-added a solution that helps the plasmid enter the cell (in addition to hitshock the next step) the
-cell suspension spent 20 microL another eppendorf and added the plasmid DNA
-the incubate 30 minutes at 42 °
-added 1 ml sterile water, centrifuged to lower-and
cells plated in the middle
To accomplish all this:
-strains grew on rich medium plates
- cells resuspended in 1 ml of sterile water and centrifuged
-discard the supernatant and resuspended in the liquid remaining
-added a solution that helps the plasmid enter the cell (in addition to hitshock the next step) the
-cell suspension spent 20 microL another eppendorf and added the plasmid DNA
-the incubate 30 minutes at 42 °
-added 1 ml sterile water, centrifuged to lower-and
cells plated in the middle
