BacterialTransformation Lab

Genetics as it applies to evolution, molecular biology, and medical aspects.

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BacterialTransformation Lab

Post by jman45 » Tue Feb 16, 2010 2:48 am

Hello everyone!

I finished my bacterial transformation lab for Bio 1 and had a few questions that I am hoping some of you can help me with. In the lab, we have 3 plates which are labeled "Transformed" containing LB media and ampicillin, "Control 1" containing LB media and ampicillin, and "Control 2" containing just LB media. The first will contain the contents of the tube containing plasmids and the EcoRI enzyme, the second contains most of the contents of the control tube (just enzyme, no plasmids), and the third plate contains 1-2 drops of the control tube. We now have to wait while the bacteria are incubated and to see whether bacteria grows on any of the plates or not. My questions are:

what can I infer if bacteria grew on the control 1 plate?
what can I infer if bateria grew on the control 2 plate?

Any help will be appreciated.. Thanks in advance

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Inland Taipan
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Post by JackBean » Tue Feb 16, 2010 7:06 am

control 1: you have non-transformed bacteria, which are able to grow in presence of ampicillin (there is high probability, that they will be also on your "Transformed" plate
control 2: you know, that your plates are OK and your cells are viable and able to grow on particular plates

BTW why did you add EcoRI to your plasmid? (just before transformation?)

Cis or trans? That's what matters.

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