I was about to design the primers for SLiCE, when I realized the insert is so small I could order it as one oligo and it would not even be that expansive.
As its quite some time ago, that I used SLiCE I became unsure, if I missed an aspect why I could not just order the oligo as an insert.
The homologues region would be 30 bp and the whole insert is 120 bp long (MW: 36977.7 [g/mol]). I can buy between 0.05 µmol and 10 µmol, that should be enough, right?
Thanks for your answer!
Best wishes
Stefanie
Hi Stefanie
maybe you'll need to design and buy a double strand DNA if you want to clone your target. Several commercial solutions are available when just typing "double strand DNA synthesis" in google.
fred
you can try to order two oligos in the small primer scale that are complementary to each other towards their 3' ends. For example, if your whole insert is 120 bp long then you can synthesize forward and reverse78-80 bp oligos from either end of the insert, having 18-20 bp complementarity to each other in the middle. Then just add the forward and reverse oligos in the PCR mixture. They will anneal and extend and generate a 120 bp primer-dimer that you can then use as your SLiCE cloning insert. The benefit is cost reduction in synthesis plus you will be able to amplify a huge amount of insert from a small scale.
Aekagra Singh Thanks this seems a very good Idea. By now, I saw that I can buy cloning oligos. I think this is exactly what I wanted to have. However, for the reasons you mention, I will consider your suggestion, too. I would end up with much more product.
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