Hi, I'm a high school student, greately inloved with genetics. I have been
working at the istitute of molecular biology here in Slovenia for about 3
years now, and I already have my own project in which I have encountered
some problems. ...Although this is not the field of microbiology, I hope
one of you had done this before :)
I'm working on diagnosis of haemophilia A. Sofar I have used only Bcl I
(in intron 18 of FVIII gene) polymorphysm for it, but now I have decited it
would be interesting to try to search for polymorphism of CA repeats
located in intron 13 of FVIII gene.
The first catch was, that I had to do two PRC reactions in order to have my
fragments (of about 140bp length) multiplied. But the real problem is, that
after electrophoresis on PAGE I se, that I actually got two strains of
about 20bp difference. (the DNA was from a man, so it couldn't have been 2
different alleles).
(I used some primers that are said to be working)
They warned me, that CA repeats are tricky, but... isn't that the thing
that makes it all fun?
So, if someone had same or similar problems with CA repeats, I would be
glad to hear them and try to avoid them :)
As far as my CA repeats are concerned, I will probably seperate the two
fragments, make another PCR and see what happens :)
...I appologise for my writing, but it's 2:45 AM now, and I'm exhausted
from the whole-day "cooking" in the lab.
Enej Kuscer
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enej.kuscer at guest.arnes.si``````~---~````````
