Cloning 180bp DNA in a vector
Pepa Florez Pérez
(by snipeurope from hotmail.com)
Sat Dec 18 16:29:04 EST 2010
I must clone a 180 bp. DNA that I have designed in several vectors.
Since it does not belong to a gene or cDNA I am not sure which one is the best method to carry out this project.
I have thought of two possibilities:
- to ask for 6 oligonucleotides, do the annealing and try to ligate them with a digested vector.
- to get the DNA synthesized by a company, and subcloning by digestion.
The problem is that I don't know which one is the proper approach, if there is any.
About the oligos, we talked about not asking them purified by HPLC or PAGE since these are quite expensive procedures, but I am afraid that becuase of that I won't have a good annealing.
And about the DNA synthesis... it cost around 200 euros/each... what we have considered as quite expensive too.
Do you think that we will be successful working with not-purified oligos? Or something cheaper will become expensive late?
Can you think of any other alternative?
Thank you very much
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