Hi~
I tried to amplify microbes' 16S rRNA gene from yoghurt sample, but I
couldn't. For this, I treated plane yoghurt with ProteinaseK during 4hr and
overnight at 37C. I found that almost proteins degraded when I centrifuged
treated sample. I washed pellet twice with PBS and once with pure water.
Finally, I added 1% Tween-80 solution and lysed cell by freeze and thaw lysis
method(I evaluated this washing and lysis method before). Lysed sample was
used directly to PCR.
I couldn't found PCR product. To evaluate sample purity, I applied sample
from yoghurt and pure culture(had used positive control) together to a PCR in
a tube. No product. I think the sample from yoghurt contains some inhibitory
materials. I don't know what material inhibits PCR. If you have some idea,
could you help me? and... If you know how can you remove them, could you tell
me that?
Adios..
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Lab. of Genetic Engineering, Inha Univ., Inchon, South Korea.
Youngmin Park bada at bigfoot.com
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