Hi all,
I am using GS Jr and its specific kits. With accurate TaqMan quantitation of libraries, I am getting the best results with DNA-to-bead ratio of 1-2 to 1. However, recently three times in a row I got 80-100% enrichment ratio even at this dpb, which happened with three different libraries. The only feasible explanation is broken emulsion, although after emPCR emulsions looked not much different than before. The yellow insert in the kit keeps mentioning that oil formulation has been changed and clear layer on the top is not indication of broken emulsion. Indeed, successful runs also had this clear layer at the emPCR step. All three kits were recent purchase, with expiration date sometime in 2013, but in all three kits I noticed unusually large precipitate, which after intence vortexing produced not "translucent", but quite turbid oil. Did anyone experience problems recently?
I am using GS Jr and its specific kits. With accurate TaqMan quantitation of libraries, I am getting the best results with DNA-to-bead ratio of 1-2 to 1. However, recently three times in a row I got 80-100% enrichment ratio even at this dpb, which happened with three different libraries. The only feasible explanation is broken emulsion, although after emPCR emulsions looked not much different than before. The yellow insert in the kit keeps mentioning that oil formulation has been changed and clear layer on the top is not indication of broken emulsion. Indeed, successful runs also had this clear layer at the emPCR step. All three kits were recent purchase, with expiration date sometime in 2013, but in all three kits I noticed unusually large precipitate, which after intence vortexing produced not "translucent", but quite turbid oil. Did anyone experience problems recently?