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Gel Electrophoresis

Gel Electrophoresis. Teacher Instructions Edvotek Set Up 12 groups. 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

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Gel Electrophoresis

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  1. Gel Electrophoresis Teacher Instructions Edvotek Set Up 12 groups

  2. 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)

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

  4. Prepare 1X SB Buffer for making gels • Pour the 50ml of 20X SB Buffer stock solution into the 1L SB Buffer mixing bottle

  5. Prepare 1X SB Buffer for making gels • 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

  6. Prepare gels • 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

  7. Prepare gels • 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

  8. Prepare gels • 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

  9. 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

  10. Prepare gels • Carefully remove the HOT bottles from the microwave and swirl - be careful of steam escaping from the loose caps!

  11. Prepare gels • Check that agar has fully dissolved or, if re-melting solidified agar, that it has all melted back into solution

  12. Prepare gels • 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

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

  14. How to store prepared gels • Each paper will hold 2 gels

  15. How to store prepared gels • Place 2 papers with 2 gels each side by side in a gel storage container • Make sure paper edges are free

  16. How to store prepared gels • 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!

  17. 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 • 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

  18. Running gels • 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

  19. Running gels • 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!

  20. Running gels • 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

  21. 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 70 • When you are ready to start the run simply press the button on the far right • Watch for bubbles in the electrophoresis boxes!

  22. After Gel Run • 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!!

  23. 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…

  24. 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

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