PCR directly from eukaryotic cell without DNA extraction

Wolfgang Schechinger hubahopp at gmx.de
Mon May 17 03:49:56 EST 2004


Dear Srinivas Vedic, 

The best source to address your problem is the library. If there is a
dedidated book on the analysis of protozoan DNA you'll be fine. If not,
find out how other people have done it before, I'm sure there are lots of
sources) with PubMed (http://www.pubmed.org) or http://www.google.com for
"protozoa DNA extraction" (e.g.). Other ideas might be to ask companies who
sell DNA exctraction kits (like Macherey-Nagel, Clontech, Sigma or Qiagen)
for help as they want to sell their stuff (and, of course, as them for free
samples before you buy).

For general methods, have a look into Maniatis, the Red Book, or Current
Protocols.

If in doubt (for science is experimental ;-) and we call it Re-Search, just
try with an ordinary DNA extraction kit or plunge your protos directly in
your PCR mix (maybe after killing them at 96degC for some time if
appropriate, and eventually inubating with RNAse. If you're lucky, it
works. If not, you'll have do do some cleanup and remove inhibitory
compounds (to find out maybe add some protos to a PCR reaction known to be
working).

Best regards,

Wo

Of course you'll meet lots of experts here in the Bionet, but it's rare
that there is knowldge available on general procedures (because nobody
likes to do the library work for you). However, if there is anything that
requires lots of experience and analytical skills (in mental sense), there
is a big chance that you will find help.



At 10:58 16.05.2004 +0100, Momba, M wrote:
>hello sir, i am srinivas vedic, from india, i am doing my phd, i want to
extract the dna from protozoans..... can i extract ?...please tell some
technique for me
>
>thanking u sir,
>
>vedic
>---
>
>
Wolfgang Schechinger


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