Sv: SAGE questions

Mette Damgaard Nielsen c929128 at student.dtu.dk
Fri Nov 17 06:02:46 EST 2000


Hi Olivier,

The reason for the two different linkers are the the PCR primer sites within
the linkers. For the PCR amplification of the ditags (as for any PCR
amplification), you need two different primers. If the primers were
identical, the two ends of the single stranded PCR would anneal, giving rise
to "panhandles", making them unable to amplify. This is actually one of the
principles in suppression subtractive hybridisation (SSH).

Good luck with SAGE !

Mette Damgaard Nielsen
Olivier Gandrillon <Gandrillon at maccgmc.univ-lyon1.fr> skrev i en
nyhedsmeddelelse:Gandrillon-EB851C.11271216112000 at news.univ-lyon1.fr...
> Hi
>
> I do have one SAGE-related question:
>
> Why are two different linkers (A and B) required? Wouldn't the all
> procedure work the same way with only one linker on both sides of the
> ditag? And in fact, aren't some ditags generated through ligating two
> tags attached to the same linker (A-tat-tag-A)?
>
>
> Thank's for your input!
>
> Olivier Gandrillon (Gandrillon at maccgmc.univ-lyon1.fr)







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