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PCR using yoghurt sample

g9711278 at inhavision.inha.ac.kr g9711278 at inhavision.inha.ac.kr
Tue Apr 14 04:13:56 EST 1998


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?


Lab. of Genetic Engineering, Inha Univ., Inchon, South Korea.
Youngmin Park   bada at bigfoot.com

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