I'm facing a problem with emulsion PCR, the emulsion phase is made of 95.05% mineral oil, 4.5% span 80, 0.4% tween 80 and 0.05% Triton x-100. For PCR, platinum pfx kit (invitrogen) is used. 50 ul of PCR was added in intervals over 2 min to 100 ul of oil mixture under stirring conditions (1000 rpm), the stirring continued for additional 5 min. then it was subjected to PCR. after PCR, i centrifuged the reaction and remove the oil phase. usually i use water saturated diethyl ether for breaking the emulsion but sometimes it does not break the emulsion completely and a creamy solution appears after the breaking step. when this happen i get very faint bands or no bands at all when migrated on agarose gel.

i tried using purification kit after this step, and i tried using isobutanol instead of water saturated diethyl ether. nothing worked.

DNA size is 90 bp