I have a transformation problem. My pet32b with insert transformed into bl21 De3 seem's to be growing to the side of the plates. What maybe the issue
1. The picture of the plate is not so clear to see. Did not see individual colonies.
2. Could be the transformed competent cells (with plasmids) did not mix well with LB medium, so when you spread them on the plate, the transformed colonies were pushed to the sides of the plates.
3. This does not mean that your transformation did not succeed. Take a few colonies from the sides of plates, grow them, miniprep them, then digest to see whether it is ok.
You can also do 'colony PCR' to check them.
Hi Sir,
Did you spread the colonies with a spreader? well if you did; while spreading this is one of the most common thing. As the spreader is repeatedly moved two and fro at the central zone of the plate while rotating the plate almost all colonies in the middle of the plate get pushed to the sides.
If 'spreading' causes a problem, you can buy a cheap petri-dish 'turntable' for this purpose. We use one in the lab. You can turn the thing while you are spreading the bacterial culture on the plate. See attachments.
This Youtube film gives you an idea of how the petri-dish turntable works. But I think the person in the film is a little bit overdo the job.
https://www.youtube.com/watch?v=3J4d7OfdOLM
1. There is also an 'electronic' version of petri-dish turntable (see attachment). But I don't think we really need a fancy one like that. Just for your information.
2. Have you checked the colonies growing on the side of the petri-dish yet? You can also re-streak those colonies (from your current plate) to the center of another new plate (with appropriate antibiotic) and see what is going to happen.
Hi tq all for the informative answer. I have actually grown a new batch of the cc. Hopefully this batch will provide the desired results
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