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DNA Structure and Analysis. Activity 4.2: Casting Agarose Gels . Activity 4.2: Casting Agarose Gels. Objectives: Calculate the amount of agarose powder needed to make 50 ml of 1% agarose Prepare gel trays for pouring the molten agarose Prepare and cast a 1% agarose TAE gel
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DNA Structure and Analysis Activity 4.2: Casting Agarose Gels
Activity 4.2: Casting Agarose Gels • Objectives: • Calculate the amount of agarose powder needed to make 50 ml of 1% agarose • Prepare gel trays for pouring the molten agarose • Prepare and cast a 1% agarose TAE gel • Remove the gel comb and store the agarose gel
Safety Reminders • Follow all laboratory safety procedures • When heating, always use heat-resistant gloves, safety glasses, and laboratory coat • Be careful when checking if agarose is completely melted. Check for superheating by gently swirling while still in microwave to ensure it will not flash boil • Be sure to use heat-resistant glass containers when melting agarose • Do not sit down when using hot liquids
Skills to Master • Prepare agarose gels
Casting an Agarose Gel • Play video: Casting an Agarose Gel
Protocol Highlights/Tips • Use 1x TAE buffer to mix with agarose to make gels • Blue painters tape works well to tape ends of gel trays • Make sure to press firmly when tape is applied to ensure proper seal • Cool agarose to approximately 55ºC so tape will not melt • If SYBR® Safe DNA stain is to be added to gel, do so just before pouring
Summary • Make sure to: • Record all steps in your notebook • Tape ends of gel tray • Seat comb well in the proper grooves in tray • Pour molten agarose so that it covers ¾ of gel comb teeth • Let cool until the agarose is well solidified • When removing gel comb, do so straight up, don’t lift one side higher than the other • If storing, leave tape on and place in a plastic bag