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oligos for ligation


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#1 vesicle

vesicle

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Posted 16 June 2010 - 04:35 AM

Hallo to everyone!

I am currently starting with a long cloning procedure.
For the beginning I have 2 oligos, 50nucleotides each, 50pmol/ul in ddH20.
I want first to anneal them according the following protocol :

200uM of each oligo
1x oligo buffer
up to 20ul ddH2O water

Questions
1): should I dilute them before in TE buffer and from that take 200uM for the reaction?
2): after annealing, should I dilute them more, or directly take from the annealed mixture something like 50nM to use in a ligation reaction with a vector?

Thank you in advance!




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