Alas, poor "magic"! I knew thee well.

Cornelius Krasel krasel at alf.biochem.mpg.de
Tue Mar 15 10:40:13 EST 1994


Martin Kennedy (mkennedy at chmeds.ac.nz) wrote:
: One point worth mentioning is that I still do 35-S manual sequencing; one of
: the people in our Lab just tried getting some sequencing of these types of
: preps done with 2 different ABI fluorescent machines, with no luck.  It seems
: additional clean up steps are necessary to get this to work.

It seems that automatic sequencing on ABI machines with Taq polymerase is
more sensitive to contamination than manual sequencing with Sequenase.
A guy in our lab brought his DNA and primers to the sequence service
(who use an ABI sequencer) and had absolutely clean lanes, no band
whatsoever; then he used the same DNA (which was, BTW, Quiagen-purified)
with a Sequenase kit from USB, and was lucky. The people who operate
the ABI machine have told us that this occurs from time to time. Has
anyone (except Martin, see above) had similar experiences?

--Cornelius.

--
/* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany */
/* email: krasel at alf.biochem.mpg.de                 fax: +49 89 8578 3795 */
/* "People are DNA's way of making more DNA." (Edward O. Wilson, 1975)    */



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