1. DNA Structure and Analysis Activity 4.2: Casting Agarose Gels

2. Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 2 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

3. Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 3 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

4. Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 4 Skills to Master  Prepare agarose gels

5. Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 5 Casting an Agarose Gel  Play video: Casting an Agarose Gel

6. Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 6 Student Workstation Materials

7. Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 7 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

8. Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 8 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