Hi Eva, if you consider the time, costs and effort to check lots of clones for the correct sequence (specially when ligating 6 unpurified oligos), I'd strongly suggest to order the whole construct, as the cost is virtually nothing (<500 USD vs. the above (and the risk of failing/ wrong results from subsequent experiments), and you have someone to blame if it does not work out. The goal of your work probably is not to establish a procedure for homemaking 180bp inserts). See eg. http://www.sloning.com and http://www.geneart.com as examples for a companies offering gene synthesis services.
HTH & good luck! Wo On Dec 18, 10:29 pm, Pepa Florez Pérez <[email protected]> wrote: > Hello everyone! > > 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 > Eva _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
