ED at micro.uct.ac.za
Mon Mar 22 05:00:53 EST 1993
> I can't help but making an analogy to help clarify my position for all
> those who wrote in and advised to never check oligo's by gel as a waste of
> If you go out to buy a half a dozen eggs, do you open the lid to see if per
> have had to do if you had purified?
> The answer is: YOU DON'T HAVE THE FOGGIEST IDEA, BECAUSE YOU'VE NEVER
> PURIFIED AN OLOGO AND DONE THE COMPARISON! But you still feel the need to
> write in and give lousy advice to someone who doesn't know better.
I didn't write in earlier, as I thought enough people would - but since
when is it a crime to offer comments based on good as opposed to bad
experience? And yes, I have purified oligos made in-house, and yes, they
were a mixture of lengthmers, and YES, THEY WORKED FINE FOR PCR - of not-
particularly-well-conserved sequence(s) from a variety of plants (verified
So - when you have a problem, check. And if you are suspicious, check.
And if you are paranoid, check. And use a program(me) such as anything
you can find if you must, but realise that someone once showed that
guesstimating oligos worked very nearly as well as using a programme, and
I found that if I had used any of the three best PC programmes available
at the time, I never would have synthesised the primers that I used
successfully to amplify the complete range of cereal geminiviruses,
because they were "unsuitable" - and I couldn't have used any other
sequences because the consensus was sufficiently high for oligo making
only in the UNSUITABLE sequences I used.
So - don't be put off from just trying 'em out anyway, un-computer
predicted, and unpurified! Having said that, I've just advised someone
with a problem to purify her oligo, because the quality control wasn't
what it should have been, but we haven't wasted any money yet!
| Ed Rybicki, PhD | "Emancipate yourselves from |
| (ed at micro.uct.ac.za) | mental slavery |
| Dept Microbiology | None but ourselves |
| University of Cape Town | Can free our minds...." |
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| 7700, South Africa | - Bob Marley |
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