First run BLASTn for the H1N1 fragments, so that you might identify the nature of the fragments. Then by using Primer3 online tool, you can design primer.

as mr Kumar said, you can use program such oligo7 or use knowen primers from articles.

Hi colleges,

Thank you for these information, but the problem with me is the designed primers aren't work so many advisers said me that the reson is the variability of the genome and others said there is unversal primers.do you have any information.

Hi Evan

so you can use papers, primers that are common in many articles often are in the best set up. you can also get a diagnostic kit from a confident company, they often work properly.

good luck

If you are still interested, you can try these set of primers: Gall A, Hoffmann B, Harder T, Grund C, Beer M (2008) Universal primer set for amplification and sequencing of HA0 cleavage sites of all influenza A viruses. Journal of clinical microbiology 46: 2561–2567.

It is working, especially if you have isolated virus in alantoic fluid (high number of genome copies).

Thank you Dejan, I'll tray it