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non-diffusion in agar

non-diffusion in agar - Protocols and Methods Forum

non-diffusion in agar - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 11-29-2005, 02:22 PM
M Des
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Default non-diffusion in agar



We have purified a small (< 1 ka) antibacterial peptide that we tested for
activity in solid media (routinely, wells cut in TSBYE, 0.75% agar, seeded
with the indicator bacteria). We also deposed a spot (10 uL) directly on the
agar. After incubation, absolutely no inhibition zone appeared around the
wells, but curiously a clear zone was visible just under the spot. A
positive control (gentamicin) was normal. Tested in liquid media, the
inoculum was totally inhibited by the peptide.

I was surprised that this antimicrobial doesn't diffuse in a soft solid
media. We supposed that the proteins agglomerate together and remained
trapped in the agar matrix, but adding Tween in the fractions did not help.
Does somebody have a clue to explain the non-diffusion of the peptide ?

Thanks

Michel
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