How can I amplify the intergenic region (IGR) of the tospo virus?

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Amanda O'Donnell

Have you tried lowering extension temperature to 60oC?

Himanshu Shekhar

Check this map may be it help you to design a proper primer.

Vishal Gurav

Thanks Amanda and Himashu for ur kind reply, I'll definitely try to follow ur suggestions.

but Earlier i've used different specific primers with various annealing temperatures. I've also used Pfu DNA polymerase but positive results r not yet found.

Lieven Gevaert

better use a pfu taq mix and or hot start pfu

http://www.gentaurpub.com/Tospovirus/VG9zcG92aXJ1cw==.html

Thank u all,

Especially Amanda, your suggestion really worked. Thanks a lot.