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dna with very low GC content?


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#1 zephyr

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Posted 27 May 2011 - 04:47 PM

Hi, I'm wondering if it is possible to create several hundred bp dsDNA composed almost entirely (90-95%) of AT?

I want to use platinum labeling (ULYSIS) to create fluorescent oligomers with very well controlled number of fluorophores.

#2 perneseblue

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Posted 28 May 2011 - 07:43 AM

I don't know. Although you could talk to a oligo synthesis company and find out.
May your PCR products be long, your protocols short and your boss on holiday

#3 phage434

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Posted 28 May 2011 - 08:06 AM

Yes, you can probably do this. The major thing to know is that standard PCR condition will likely not work, due to the melting temperature of the 20 bp or so region at the 5' end of the extending strand being lower than the extension temperature used during PCR cycling. You must lower the extension temperature (not the annealing temperature, although that might need to be low as well) to 62-66C rather than the standard 72C.




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