File:MIT2012 Rescuing Loss of Flurescence by Adding Excess Carrier RNA medium.png

Effect of carrier RNA on fluorescence levels. Previously () it has been shown that RNA and DNA stick to plastics in a non-specific way. This includes pipette tips as well as wells in a plate reader plate. Thus, for accurate measurements, a carrier of some sort is needed, otherwise a decrease in fluorescent signal is observed as the nucleic acids sticks to the sides of a well.
Here, Part:BBa_K779119 is used as an excess carrier (>10 uM whereas dsROX was at 40 nM). Measurements carried out on a Tecan Safire II plate reader (see [http://2012.igem.org/Team:MIT/MaterialsAndMethods#iv2bio protocol]) using a 384-well plate. At timepoint 1, the wells are mixed using a pipette, disturbing the RNA bound to the well. At timepoint 2, carrier RNA is added, which now outcompetes dsROX for binding to the plate. Almost all of the fluorescence is recovered.