You should be able to do this using any site-directed mutagenesis methods. For example look up the Quick Change method.

Thank you Michael, I will go for this approach.

Try this way:

1. design primers to amplify the entire vector (but those 15 bp) = you will delete them.

(use Extreme KOD hot start DNA pol . or other proofreading)

2. During amplification first use only one of the primers and run few cycles;

3. Then add the other primer and run 30x cycles.

4. Purify the PCR product; (here is important that you phosphorylate the product if you do not use 5’ primers. You can use also the NEB blunting kit (15-30 min) and it has kinase activity…or use PNK

5. self ligate and you done

GOOD LUCK

I mean > 5'phosphorilated primers

Thank you Zoia!

You can do that by means of restriction free cloning approach. It is similar as site directed mutagenesis and you can delete more base pairs. For this you have to design the primer which does not contain the base pairs(15bp) which you want to delete from your gene.For better and detail explanation have a look through this paper.

http://ftp2.mrc-lmb.cam.ac.uk/groups/JYL/PDF/rfcloning2006%20published.pdf

You have to treat you PCR reaction with DpnI to remove the plasmid DNA used as template. You can do that after the PCR amplification. Sorry I forgot to mention that

I think the Shiva has the best suggestion. Overlapping extension PCR s the quickest method. My colleagues use it frequently. You can read this for starters http://en.wikipedia.org/wiki/Overlap_extension_polymerase_chain_reaction. Or just type into Google and there are many diagrams. I works really well and it can all be done in one day. As long as you use decent taq then no mutations should occur either. Hope this helps

I recently purchased the NEB Site directed mutagenesis kit, which looks quite straightforward and is inexpensive ($180). However, I have not tried it yet.

I recently purchased the NEB Site directed mutagenesis kit, which looks quite straightforward and is inexpensive ($180). However, I have not tried it yet.

https://www.neb.com/products/e0554-q5-site-directed-mutagenesis-kit

Thanks for all the suggestions!