Gel Electrophoresis Teacher Instructions Bio Rad Set Up

Gel Electrophoresis Teacher Instructions Bio. Rad Set Up 8 groups Grossmont Kit Salk Institute Mobile Lab

Timeline • Prepare Gels: Up to a week in advance • Class lab: 1 -3 days – Teach students to pipette – Load and run gels – Teach electrophoresis theory – Analyze gels • Gels must be analyzed no later than next day after running (stored in refrigerator overnight) Salk Institute Mobile Lab

Prepare 1 X TAE Buffer for making gels • Measure 28 ml of 50 X TAE Buffer stock solution into the 50 ml conical TAE Buffer measuring tube Salk Institute Mobile Lab

Prepare 1 X TAE Buffer for making gels • Pour the 28 ml of 50 X TAE Buffer stock solution into the 2 L TAE Buffer mixing bottle Salk Institute Mobile Lab

Prepare 1 X TAE Buffer for making gels • Fill 2 L TAE Buffer mixing bottle to the 1400 ml line with water (tap or distilled) • You might need to repeat this as necessary for your number of classes – this bottle should prepare enough 1 X TAE Buffer for 6 classes worth of gels Salk Institute Mobile Lab

Prepare Agar for Gels • Measure 3 level scoops of agar powder into each glass agar mixing bottle – Each scoop is 1/4 tsp – This will equal ~1. 5 g of powder per bottle • You’ll need to make 1 bottle per class Salk Institute Mobile Lab

Prepare Agar for Gels • Fill each bottle to the 200 ml mark with your prepared 1 X TAE Buffer solution – Bottles have been prechecked for calibration • Cap tightly and shake to mix Salk Institute Mobile Lab

Prepare Agar for Gels • Loosen caps slightly and place bottles in your microwave • Set microwave for 1 -2 minutes per bottle (less is better - you can always add more time!) • Allow agar solution to come to a boil - stop microwave once a good boil starts Salk Institute Mobile Lab

Prepare gels • Carefully remove the HOT bottle from the microwave and swirl - be careful of steam escaping from the loose caps! Salk Institute Mobile Lab

Prepare Agar for Gels • Check that agar has fully dissolved or, if re-melting solidified agar, that it has all melted back into solution • Allow Agar to cool until you can just stand holding the bottle with your bare hand Salk Institute Mobile Lab

Prepare Gel Casting Trays • While Agar is cooling use blue lab tape to carefully seal the end of each tray • Place 2 combs into slots on each tray (top and middle) Salk Institute Mobile Lab

Pour Gels • Carefully pour warm agar solution into the assembled gel casting trays (make sure agar is still fully melted) • Fill each tray approximately. 75 cm • 1 bottle should fill 4 trays Salk Institute Mobile Lab

How to store prepared gels • After gels have solidified, remove the combs and tape • Carefully slide gel out of the tray onto a “patty pac” paper • 1 gel per paper • Re-tape tray to pour for next class set Salk Institute Mobile Lab

How to store prepared gels • Place 2 papers with 1 gel each side by side in a gel storage container • Make sure paper edges are free Salk Institute Mobile Lab

How to store prepared gels • Stack second layer of gels in storage container and place container in fridge for up to a week • 1 container per class! – (4 full size gels) Salk Institute Mobile Lab

Setting up prepared gels for class • When you are ready to have students use gels simply carefully lift paper with gels out of the storage container • Carefully use spatula to cut each gel in half (~1 cm above middle wells) Salk Institute Mobile Lab

Setting up prepared gels for class • Place each gel half onto flat tray for each student group – Put a patty pac under if too hard to see wells • Try to keep gels and trays low and level to prevent accidental tearing of the gel Salk Institute Mobile Lab

Running gels • Prepare 1 X TAE Buffer solution as before: – Pour the 28 ml of 50 X TAE Buffer stock solution into the 2 L TAE Buffer mixing bottle – Fill 2 L TAE Buffer mixing bottle to the 1400 ml line with water (tap or distilled) • Pour 1 X TAE into electrophoresis boxes stopping about 1 cm below MAX line Salk Institute Mobile Lab

Running gels • After students have loaded their gels carefully slide each gel half back into each gel tray (in same direction) and place into the electrophoresis boxes – Keep track of which groups’ gels are where! • Make sure the well sides of the gels are on the BLACK electrode side – Back to Black, RUN to RED Salk Institute Mobile Lab

Running gels • Top off each box as needed with TAE to bring level to MAX line • Place lid on each electrophoresis box making sure that the black electrode is at the well end of the gels Salk Institute Mobile Lab

Running gels • Connect the electrodes from each box to the power supply and turn on the power by the switch in the back • Make sure the power supply is set on volts and adjust the voltage to 200* • When you are ready to start the run simply press the button in the center – Watch for bubbles in the electrophoresis boxes! *may need to lower voltage Salk Institute Mobile Lab

After Gel Run • When the run is complete (colors have separated) turn off the power by pressing the button on the left • Remove the lids from the electrophoresis boxes Salk Institute Mobile Lab

After Gel Run • Carefully remove gel trays from the box and depending on time: – Place each ½ gel on a flat tray on a patty pac and give back to groups to analyze – OR place each ½ gel on a labeled patty pac and store back in storage container in refrigerator until next class meeting and then distribute on flat trays • WARNING - WET GELS ARE VERY SLIPPERY!! Salk Institute Mobile Lab

Next period and so on… • You can prepare per 2 gels for distribution while per 1 gels are running and so on… • Running TAE buffer is good for all classes – no need to replace unless it gets too hot Salk Institute Mobile Lab

Clean up • At end of day used buffer can just be flushed down sink – Rinse boxes and let air dry • Used gels can be placed in general trash Salk Institute Mobile Lab