Salk Institute Mobile Lab Gel Electrophoresis Teacher Instructions Edvotek Set Up 12 groups

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 1X SB Buffer for making gels Measure 50ml of 20X SB Buffer stock solution into the 50ml conical SB Buffer measuring tube

Salk Institute Mobile Lab Pour the 50ml of 20X SB Buffer stock solution into the 1L SB Buffer mixing bottle Prepare 1X SB Buffer for making gels

Salk Institute Mobile Lab Fill 1L SB Buffer mixing bottle to the 1000ml line with water (tap or distilled) You’ll need to repeat this as necessary for your number of classes - each bottle should prepare enough 1X SB Buffer for 2 1/2 classes Prepare 1X SB Buffer for making gels

Salk Institute Mobile Lab Measure 3 level scoops of agar powder into each glass agar mixing bottle –Each scoop is ~1/4 tsp –This will equal ~1.5g of powder per bottle You’ll need 2 bottles per class Prepare gels

Salk Institute Mobile Lab Fill each bottle to the 200ml mark with your prepared 1X SB Buffer solution –Bottles have been pre- checked for calibration Cap tightly and shake to mix Prepare gels

Salk Institute Mobile Lab Loosen cap 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 full boil starts Prepare gels

Salk Institute Mobile Lab Prepare gels While agar is in microwave assemble the gel trays –Can assemble 24 trays at a time Place rubber gaskets on each side of the gel casting tray Place comb in end slots

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

Salk Institute Mobile Lab Check that agar has fully dissolved or, if re-melting solidified agar, that it has all melted back into solution Prepare gels

Salk Institute Mobile Lab Carefully pour hot agar solution into the assembled gel casting trays (okay to pour while really hot) Fill each tray to top of rubber gaskets Each bottle should fill 6 trays Prepare gels

Salk Institute Mobile Lab After gels have solidified, remove the comb and both gaskets Carefully slide gel out of the tray onto a “patty pac” paper How to store prepared gels

Salk Institute Mobile Lab Each paper will hold 2 gels How to store prepared gels

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

Salk Institute Mobile Lab Stack rest of gels in storage container and place container in fridge for up to a week Each container will hold 12 gels - 1 container per class! How to store prepared gels

Salk Institute Mobile Lab When you are ready to have students use gels simply carefully lift paper with gels out of the storage container Then gently slide each gel back into a casting tray –Be sure to slide gels into trays in same orientation they were cast - (wells at notch end of tray) Try to keep gels and trays low and level to prevent accidental tearing of the gel Setting up prepared gels for class

Salk Institute Mobile Lab Prepare 1X SB Buffer solution as before You’ll need 2 L of solution total for both electrophoresis boxes Place tray supports in each electrophoresis box and pour in 1L of prepared 1X SB Buffer into each box You only need to do this before the first class Running gels

Salk Institute Mobile Lab After students have loaded their gels carefully place each gel tray into the electrophoresis boxes The trays will only fit in one direction ;) Remember to use care that the gels don’t slide out of the trays as they are carried to the boxes! Running gels

Salk Institute Mobile Lab Place lid on each electrophoresis box making sure that the black electrode is at the well end of the gels –The electrode wires inside the box are color coded as well Running gels

Salk Institute Mobile Lab 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 70 When you are ready to start the run simply press the button on the far right –Watch for bubbles in the electrophoresis boxes! Running gels

Salk Institute Mobile Lab After the colors have separated turn off the power supply and remove the gel box lid Carefully remove gel trays from the box and depending on time: –Give back to groups to analyze –OR place each gel on a labeled patty pac and store back in storage container in refrigerator until next class meeting and then distribute on individual patty pacs WARNING - WET GELS ARE VERY SLIPPERY!! After Gel Run

Salk Institute Mobile Lab Next period and so on… Running SB buffer is good for all classes – no need to replace unless it gets too hot You can put gels into trays for periods 1 and 2, then after removing completed gels from per 1 trays put gels for per 3 into trays while per 2 is running and so on…

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