FRET - Slide Preparation (Page 2)
We keep stocks of 1% SeaKem Gold Agarose (for lower background in the CFP channel and high tensile strength) prepared in S media (1.7 g yeast nitrogen base w/o amino acids and 5 g ammonium sulfate in 900 ml) in 0.45 ml aliquots in 1.5 ml eppendorf tubes stored in our refrigerator. To prepare the pads a tube of agarose is melted in boiling water, and then equilibrated to 65 C. We prepare a fresh mix of 10X nutrients by combining 500 ul of 40% glucose, 100 ul of 5 mg/ml adenine, 100 ul of 2.5 mg/ml uracil, 100 ul of filter sterilized 10% casamino acids and 100 ul of 1% filter sterilized tryptophan (stored refrigerated). To the tube of agarose is added 50 ul of the 10X nutrients to make the standard SDC medium.
A 30 ul aliquot of the SDC agarose is pipetted into the center of the microscope slide.
