I have designed 5'phosphorylated linker with sticky end and restriction sites. I have 25 nmol conc of each separate strand.
Here is the sample construct:
strand 1.
5 ....agctt---------18bp----------gcggccgcac... 3
HindIII site 3.............a---------18bp-------gccggcgtgagct....5 xhoI site
I would really appreciate if you could tell me the following:
1. What should the solvent be for these linkers? Water or some buffer?
2. How should I anneal these strands together?
3. What should be the appropriate ratio of double digested vector and annealed linker?
4. Is concatemer formation possible in this case?
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