PCR using yoghurt sample

g9711278 at inhavision.inha.ac.kr g9711278 at inhavision.inha.ac.kr
Fri Apr 10 03:10:04 EST 1998


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..

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

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