Annealing oligos for cloning?

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Lenin Yong

This protocol worked fine for me:

http://www.sigmaaldrich.com/technical-documents/protocols/biology/annealing-oligos.html

As for the ligation i usually calculate the dsDNA amount as for normal ligations, molar ratios 1:1 and 1:10 are optimal for single insertions (up to 1:20 for short adaptors).

Norbert Duró

Thanks for the answers David and Lenin!