I have designed the primers for quick change mutagenesis in agilent software....I checked the heterodimer, homodimer and hairpin in one oligo analyzer...http://eu.idtdna.com/analyzer/applications/oligoanalyzer/. In that i found out that the primers had high energy to form homo and heterodimers (< -3 kcal/mole) i.e. both my primers have -9 kcal/mole and -6 kcal/mole energy. So, i looked for better primers, by reducing the bps but it too resulted similarly. Now what shall i do? Already the primers which i used did not work so im designing new one...So plz help me out in getting good primers.Also comment on the primers i have designed...Waiting for your reply...Plz do reply soon
