Materials

• 1X TBE
• Ethidium bromide
• DNA ladder
• PCR reactions
• Gel electrophoresis system
• Agarose

Recipes

20X Sodium borate buffer (SB)

8 g NaOH pellets

48 g boric acid

900 mL deionized water

Adjust pH to 8

Deionized water to 1 L

Add 500 mL to 9.5 L deionized water int he carboy

Setting up the gel

1. Make 50 mL of a 1% agarose solution in TBE
1. 50 mL buffer
2. 0.5 g agarose
3. Heat in microwave until dissolved (5-10 sec at a time)
2. Let cool for 2-3 minutes
3. Add 5 µL ethidium bromide
4. Pour gel into cast (with comb)
5. Let polymerize (10-15 minutes)
6. Add to gel box
1. turn casting tray 90°, wells on side with negative electrode
7. Cover with 1X TBE (from used beaker)
8. Load DNA ladder and PCR reactions
1. 10 µL ladder (first well)
2. 5 µL PCR + 1 µL loading dye
9. Plug power cables into corresponding electrodes
10. Run gel at 80 V until bands are fully expanded

Imaging the gel

1. Use the imaging system
1. open imaging app (select Nucleic Acid Gel, E Bromide filter)
2. place gel in on tray, close
3. UV light
4. live view
5. adjust focus/ brightness
6. take image
2. Crop the image
3. Name the image: date_initials_number if more than one in a day
1. ex/ 20230607_MRB_2
4. Transfer images to OneDrive using the USB drive labeled WHEELER LAB GELS (the USB drive can be found in P-366 drawer M-11)
5. Dispose of the gel in biohazard container
6. Drain TBE into used flask