I'm doing phage display recently. And I am curious about why many blue plaques on my IPTG/Xgal plates have little colorless area in the center. This phenomenon had not happened before the third round titering. Would it be possible that my phage lose specificty? My friends also have the same problem; However we couldn't figure out why. Please tell us if you observed the same things as we did. And maybe we can discuss together. Thanks for every advanced answers.
i would rather think of dieing cells in the middle, and loss of colour during this.
Hi Chih-Chun,
Your phage should increase in specificity towards your target as you increase the number of rounds of panning. The color in the middle shouldn't be an indication of loss of specificity. Loss of specificity can occur if you perform intermediate amplification in your rounds of panning. I avoid amplifying my phage until my 3rd round of biopanning is complete.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Biological Techniques