Multigene cloning - Looking for a cheap solution

[vbbgold]

Hi there,

I need to insert 5 DNA fragments into 1 vector. It is impossible to find restriction sites to do this.

I found Gateway from Invitrogen is a good solution. However, it is too expensive.

Any suggestion for a cheaper solution?

Thanks heap!

[snowcapk]

What do you mean by 5 fragments in one vector? Do you want (a) to clone each fragment separately, or do you want (b) to insert all five of them in a row in a single vector?

(a) Primers are very cheap. Why don't you just design ten primers with rare restriction sites in the tails, amplify your five fragments with them, then digest with the appropriate restriciton enzymes? Any vector your lab already has could be acceptable then.
(b) Google fusion PCR.