To troubleshoot this issue:
- Verify the contents and sequence of your plasmids to check if they contain any red markers.
- Double-check the strain and plasmid you are using to ensure there are no mix-ups.
- Repeat the transformation experiment with controls to assess the transformation efficiency and check for unwanted genetic changes.
- Consider performing a genetic analysis, such as PCR or sequencing, to confirm the presence and location of the plasmid in the transformed cells.
If you continue to encounter issues, it may be helpful to consult with colleagues or experts in the field or seek assistance from your laboratory supervisor to troubleshoot the problem further.
Are you sure you have enough Adenine? You might try a different Adenine stock solution (unless you have successfully used yours in the past) and / or maybe a new batch of plates.
I agree with Mchael J Benedik, your Adenine stock may be insufficient, may have expired, or your plates might be the problem. Did you make the plates yourself? Anyone else using the same plates and seeing the same phenotype? Can you streak/plate another strain on the same batch of plates to see if it is specific to your transformation or a more general occurrence?
The red color by itself isn't necessarily a problem. You might still have what you want.
My colleagues are facing the same problem. And the plates cannot be the problem since in another experiment I have used the same batch of plates for spotting.
Hmm, was the plate spotting at the same time as the transformations? Medium in plates can break down over time.
My first thought would be to make new plates, it's an easy thing to try.
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