Medium and Buffers

20% (w/v) glucose solution

1. Prepare the solution by mixing the ingredients stated above.
2. Sterilize in an autoclave before using it to prepare the SOC medium.

50% (w/v) glycerol solution

1. Prepare the solution by mixing the ingredients stated above.
2. Sterilize in an autoclave.

*Source from 2015 iGEM Exeter [link = 2015.igem2015 .org/wiki/images/a/ab/Exeter_Glycerol_recipe.pdf]

Electrophoresis buffers

For nucleic acid DNA/RNA separation

• TAE buffer (Tris-acetate-EDTA)
• TBE buffer (Tris-borate-EDTA)
• LB buffer (Lithium borate)

1. Prepare the solution by mixing the ingredients stated above.

LB agar (Luria-Bertani agar)

If not using pre-mixed LB agar powder, prepare the materials as below:

1. In a 1L Erlenmeyer flask, swirl and mix the solution.
2. Cover the top of the flask with a lid/aluminum foil and label with autoclave tape.
3. Autoclave the liquid setting for 20 minutes or according to your autoclave's specifications.
4. After removing the solution from the autoclave, allow the agar solution to cool to 55°C in an oven or water bath.
5. When pouring the LB agar into plates, keep the bench area sterile by working near a flame or Bunsen burner. Alternatively, prepare the plates in a vacuum hood.
6. Add the appropriate amount of desired antibiotic (refer the table below) to the solution and swirl to mix.
7. Pour approximately 20mL of LB agar per 10cm polystyrene Petri dish.
8. Place the lids on the plates and allow them to cool for until the agar is solidified.
9. Label the bottom of plates with antibiotic and date before storing in plastic bags or sealed with Parafilm at 4°C.