I am using M13 phage peptide library for biapanning of specific peptide. After first 2 rounds of screening, i am now observing a blue layer on agar plate. I have diluted the phage upto highest dilution but still having the same problem from lowest to highest dilution? What might be the problem ? How to overcome it ?
If you are not using an Xgal selection plates I suggest to check your library, probably you have some contaminations.
What host strain are you using to amplify your phage? If the strain is lac+ then you may just have residual B-gal activity from the cultures.
Thanks for the suggestion. I have overcomed it after drying the IPTG/Xgal agar plate for few more hours before plating phage mixed bacteria and also using fresh bacterial culture from the stock.
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