These isoforms are highly homologous in the center of the cDNAs, so i can not look for primers in these homologous regions, right?
so i tried in the 5' end and the 3' end(i don't know if it is right to look for primers in the 3' UTR region of cDNAs, can you tell me ?).But there are soooooo many repeats of bases in these two regions that whatever primers i picked up had seemed to be unspecific when do BLAST with Arabidopsis genome(usua. there were 10 unspecific pairings )
the software i uesed:Primer3 online; Tair PCR;
the parameters i used : Tm:62-64 degree centigrade; product length:100-200bp; primer length:22-24bp;
I really need help.....
Edited by bear, 06 March 2009 - 08:06 PM.