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[[Media:Agarose Gel_Synthesis_Protocol.pdf|Agarose_Gel_Synthesis_Protocol PDF]] ==Agarose Gel Synthesis Protocol == ==Overview== *Determine the concentration of agarose to be used *Measurement of agarose gel and TBE buffer *Mix and Heat *Cool Down *Gel Staining *Casting *Storage ==Samples & Material== *Erlenmeyer Flask (250mL) *Graduated Cylinder *Mircowave *Gel tray 13 x 12.2cm (2 standard gel trays) *Gel Tray (4 micro gel trays) *Combs for micro gels (6/8 teeth) *Standard combs (13/26 teeth) *Gel casting stand for micro gels *Gel casting stand *0.5X TBE Buffer *Ultra Pure Agarose -Invitrogen Cat:15510-027 *SYBR Safe DNA gel stain10,000X concentration in DMSO -Invitrogen Cat:S33102 ==Protocol== *Determine the concentration of Agarose to be used (1 Tray) DNA Size 5 - 10 kb 0.2 - 1 kb Concentration of Agarose 0.7 % 2 % Amount of Agarose 0.7 g 2 g Amount of 0.5X TBE Buffer 100 mL 100 mL Amount of SYBR Safe 8uL 8uL Note: Scale up or down accordingly *Weigh out the required amount of Agarose with a scale *Measure the required volume of TBE buffer using a graduated cylinder Caution: Hot Agarose solution should be handled very carefully. *Transfer the Agarose powder into a 250mL Erlenmeyer flask and add in the TBE buffer, mix by swirling the flask and microwave for 30s *Repeat mixing and heating step until all gel powder are dissolved and the solution appears to be clear *Let cool for a while and add in the required amount of SYBR Safe, swirl flask to mix the dye *Cast gel into tray with the required comb set in the tray *Let cool
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