gottainvent
Posts: 4
Joined: Tue Aug 14, 2007 11:36 am

PGreen Plasmid kit transformation help

Postby gottainvent » Tue Aug 14, 2007 11:45 am

Hi guys, :arrow:
I am doing a project on Bacterial transformation. It is a kit I bought from Carolina. It is the PGreen Plasmid, to make the transformed cells glow green. I did the experiment, but the result could not be seen. There were colonies but they did not "glow" under UV light, just faint colour. But as there were colonies the plasmid had been taken up because the medium had ampicillin and the plasmid also had plasmid for amp resistence. So I was wondering if anybody could help me to determine what I did wrong? I did start on the kit late could that be a problem? But I am sure I did the transformation correctly. 8) :?:

Louise
Former Expert
Posts: 921
Joined: Mon Jan 16, 2006 2:17 pm

Re: PGreen Plasmid kit transformation help

Postby Louise » Tue Aug 14, 2007 12:54 pm

gottainvent wrote:Hi guys, :arrow:
I am doing a project on Bacterial transformation. It is a kit I bought from Carolina. It is the PGreen Plasmid, to make the transformed cells glow green. I did the experiment, but the result could not be seen. There were colonies but they did not "glow" under UV light, just faint colour. But as there were colonies the plasmid had been taken up because the medium had ampicillin and the plasmid also had plasmid for amp resistence. So I was wondering if anybody could help me to determine what I did wrong? I did start on the kit late could that be a problem? But I am sure I did the transformation correctly. 8) :?:


I would call Carolina and ask them. Also, I'm not sure how bright you would expect it to be. If you saw faint color= green, then maybe that is it.

The contact info is:
"If you have questions about any of our products, or need help selecting or using materials for specific applications, call our professional staff at 800-227-1150 (8 a.m. to 5 p.m., ET, M–F)"


Louise

gottainvent
Posts: 4
Joined: Tue Aug 14, 2007 11:36 am

Postby gottainvent » Thu Aug 16, 2007 8:20 pm

Thanks 8) I'll try it

Louise
Former Expert
Posts: 921
Joined: Mon Jan 16, 2006 2:17 pm

Postby Louise » Mon Aug 20, 2007 11:50 am

gottainvent wrote:Thanks 8) I'll try it


Hey, when you post again, could you either link to the instructions you used, or paste your procedure here? Also, if you did any controls, did they work okay? (how many colonies did you get for the control? how many colonies grew in the amp. plate (and only one was glowing?)? how many plates did you do of each? etc.)

Here is an example of the type of detail needed (and probably the level of detail you should be giving carolina when you call):

http://www.sciencebuddies.com/mentoring/project_ideas/BioChem_p013.shtml?from=Home


Louise


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