Cloning by multiple oligo ligation
Posted 02 October 2009 - 08:09 AM
1) Synthesize a series of + and - sense oligos that anneal to each other, such that they create a (nicked) dsDNA fragment encoding my peptide with unique restriction sites at each end
2) Use ligase to join the nicked dsDNA into a continuous fragment
3) Ligate into expression vector
The modular nature should allow me to make mutations in my peptide by simply swapping out a pair of oligos. If anyone has experience with something like this method, I'd love to hear your opinions, tips, pitfalls to watch out for, etc.
Posted 02 October 2009 - 02:54 PM
Posted 03 October 2009 - 03:02 AM
Yes. It is mandatory for gene synthesis!