I'm glad to announce the trial was succesful .... :) 4.5 kb :) without noise

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC310446/pdf/nar00077-0233.pdf

Gracias!

yes you are in the right point 2 -3 kb is perfect for a regular TaqPol my fragment is in 5 Kb, using a platinum polimerase and Phusion Polimerase for that reason im searching literature in internet i've found the use of the mixture of Taq and Pfu pol but is not what i'm looking for, but thanks i will search using different ways in the advance search regards

Probably you've tried it by now, but if not the suggestion above to increase time, especially the polymerization step is very good one, the rule of thumb is 1min per 1kb, but may need additional optimization. Also decrease the pol. temp. to 68 degr.

There's no need to buy a specific kit to amplify 3Kb amplicons. We routinely use Phusion Taq to amplify plasmids of up to 10Kb. The amplification works perfectly at 15 seconds per Kb. That's 4 times faster than a typical Taq and 8 times fatser than Pfu

We have amplified ~5 kb fragments with Phusion without any special modifications. Estimate 30 sec/kb for extension. You really want to use a high-fidelity enzyme like Phusion as the error rate will become significant with lower-fidelity enzymes like Taq.

I'm glad to announce the trial was succesful .... :) 4.5 kb :) without noise

Hi Camilo. Could you please share the protocol that worked for you