Building genes with oligos

Benoit_Hebert at IAF.UQUEBEC.CA Benoit_Hebert at IAF.UQUEBEC.CA
Tue Jul 19 20:32:44 EST 1994


glenn soltes (gsoltes at gpu.utcc.utoronto.ca) wrote:
> Glenn Soltes
> Cangene Corp
> Our lab has had difficulties building genes from
> complementary 40-60 nt oligonucleotides generated by the
> Applied Biosystems 392 DNA/RNA synthesizer.
...
> Does anyone know what is causing the mutations?
> Is it:
>  a) Probabilistic errors in the oligo synthesis.
>  b) UV exposure mutations (during UV shadowing).
>  c) Secondary structure of the oligos.
>  d) Recombination/repair

If you are concerned with errors incorporated in your oligos during synthesis, 
you may want to look at the pattern of "mutation". That is, do you always get 
Cs instead of Gs? Although I'm not familiar with this synthesizer, I've had 
problems with oligos from a Pharmacia Gene Assembler. The problem came from a 
leaky rotary valve. The nu being under positive nitrogen pressure, leaks occur 
easily. After changing the valve, no more errors... Just a though.

Good luck!

Benoit Hebert

========================================
Benoit_Hebert at iaf.uquebec.ca

Virology Research Center
Armand-Frappier Institute, Laval, Quebec
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