T-top (0.5% agar)-PDF

Materials

500 mL:

• 5 g Tryptone
• 5 g NaCl
• 500 ml dH₂O
• Bacto agar

1000 mL:

• 10 g Tryptone
• 10 g NaCl
• 1000 ml dH₂O
• Bacto agar

Procedure

Dissolve tryptone and NaCl in water. Add Bacto Agar to square milk bottles to a final concentration of 0.5 % w/v (0.25 g / 50 ml, 0.5 g / 100 ml). Aliquot medium to bottles as appropriate (50 or 100 ml), autoclave and store at RT. To use, melt an aliquot of top agar in a microwave and add MgSO₄ and CaCl₂ from sterile 1 M stocks to a final concentration of 5 mM each.

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!