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 :
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
just to correct a point in your awesome french (très bien écrit d'ailleurs )
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.