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Annealing of oligos - (Dec/07/2005 )


I want to prepare an adapter for blunt end ligation.
First strand of the adapter is ~ 40-50 nucleotodes. The second strand is 5' and 3' modified and is ~ 8 nucleotides long.
As for the huge difference between these strands (just ~ 8 bases complementarity) is there anything I should be aware of or could anybody recommend a good protocol for annealing?
Thanks all of you for giving input in advance!


big difference in these two sizes.
Btw : i posted my protocol for annealing here :

that topic may help too :


Salut Fred!

Merci beaucoup pour ton repondre. Je vais essayer les protocols!

Hoho, just kidding: thanks for your answer. Do you think I can proof succesful annealing by a simple gel?

I want to try a protocol with ligation, PCR and nested PCR. Would be really f*#king annoying if the annaeling step already fails...and I couldn´t proof it... .
Thanks a lot!

A plus tard cool.gif


just to correct a point in your awesome french (très bien écrit d'ailleurs laugh.gif )
we actually write "protocoles".
and repondre is written répondre and you meant réponse...
ah it's a crazy language...

have a nice day and succesful exp.