DNA Structure and Analysis Activity 4.2: Casting Agarose Gels.

DNA Structure and Analysis

Activity 4.2: Casting Agarose Gels

Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com2

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


Student Workstation Materials


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

DNA Structure and Analysis Activity 4.2: Casting Agarose Gels.

Documents

Transcript of DNA Structure and Analysis Activity 4.2: Casting Agarose Gels.