Hello,
I have currently been tasked with performing a split GFP analysis.
I have been using DH5a cells for plasmid transformation. The results were turning out to be what we expected. The strains hosting just one half of GFP did not fluoresce while the strains hosting the reconstituted GFP did fluoresce. All strains were induced with arabinose.
However, I ran into an issue. I decided to grow up the parent cell strain in an overnight culture and introduce them to arabinose. This is to ensure that the parent cell did not fluoresce without the plasmids. (I just took some cells from the competent cell strain tube that I used to do the split-GFP transformations)
The results showed that DH5a did not fluoresce when it was introduced to arabinose. However, DH5a did fluoresce when it was not introduced to arabinose
So, I decided to test if the MC4100 strain fluoresced, and it did indeed. MC4100 fluoresced with or without arabinose.
I am wondering if anyone has any possible explanation for this phenomenon? I know E. Coli has some background fluorescence, but I had no idea that the DH5a variant was affected by arabinose.
Is it possible that they are fluorescing because they were grown up from competent cells?
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