addressalign-toparrow-leftarrow-rightbackbellblockcalendarcameraccwcheckchevron-downchevron-leftchevron-rightchevron-small-downchevron-small-leftchevron-small-rightchevron-small-upchevron-upcircle-with-checkcircle-with-crosscircle-with-pluscrossdots-three-verticaleditemptyheartexporteye-with-lineeyefacebookfolderfullheartglobegmailgooglegroupshelp-with-circleimageimagesinstagramlinklocation-pinm-swarmSearchmailmessagesminusmoremuplabelShape 3 + Rectangle 1ShapeoutlookpersonJoin Group on CardStartprice-ribbonShapeShapeShapeShapeImported LayersImported LayersImported Layersshieldstartickettrashtriangle-downtriangle-uptwitteruserwarningyahoo

BioCurious Message Board › GFP Class Results

GFP Class Results

Paul J.
user 13392631
Sydney, AU
Thanks everyone for such a great class, I had a lot of fun teaching you how to transform plasmids into E. coli. The results are in, and I'm happy to say it was a complete success! Everybody got a lot of transformant colonies on their plates.

Photos of plates:




Diane & David­

Dan & Andrew­

Sydney S.­



David & Nick­

Patrick, Adam, and Phil­



Some strange results I saw included glowing E. coli colonies on the lid of Sydney S. group's amp+ara plate and somehow Fitsoom's 30 min amp plate had glowing on one corner without arabinose!
You can see that some of the amp+ara plates look quite green under normal white light, showing just how much GFP they are expressing.

Also, some groups had glowing colonies on their ara-only plates. By comparing the white light photo to the black light photo you can see the very tiny proportion of cells that actually get the plasmid.

Prize for the most number of GFP transformants would probably go to Sydney S. although most had very similar amounts. Artistic award goes to the RT LL group for writing initials that worked out pretty well.

For those of you interested in additional details of the experiment, the E. coli strain we used was BW27783 (http://cgsc.biology.y...­, which has an arabinose membrane transporter (a protein in the membrane that specifically transports arabinose into the cell) introduced onto the chromosome so that the response of the cell to arabinose is linear rather than the on/off behavior seen in cells without the transporter. The strain was made 10 years ago by the Keasling lab at UC Berkeley.

We used 0.5% arabinose in the plates (which is the high range of what you'd want to use. usually 0.1 - 0.5% (mass to volume) is used). Ampicillin was at 150 µg/mL.

Until next time,
Powered by mvnForum

Our Sponsors

People in this
Meetup are also in:

Sign up

Meetup members, Log in

By clicking "Sign up" or "Sign up using Facebook", you confirm that you accept our Terms of Service & Privacy Policy