User:David.lim.yale/5 July 2012

From 2012.igem.org

Make 10X phosphate buffered saline (PBS) stock solution

• For 1 L of 10X PBS (pH ~6.8 before dilution to 1X), mix:
  • 80 g (80.06) NaCl
  • 2 g (1.99735) KCl
  • 14.4 g (14.42360) Na₂HPO₄ (anhydrous)
  • 2.4 g (2.39191) KH₂PO₄ (anhydrous)
• Add dH₂O to 1 L, mix to dissolve solids
• Autoclave solution
• To dilute to 1X PBS (pH 7.2-7.4), add 900 mL sterile H₂O to 100 mL of 10X PBS stock solution

Make 5 mg/mL Tet stock solution for plates/liquid cultures

Note: tetracycline is light-sensitive, so all protocols involving it must be carried out in the dark

• Weigh out 100 mg (99.87) of Tet powder from refrigerator
• Bring to 20 mL with absolute EtOH, mix gently to dissolve solids
• Aliquot into foil-covered Eppendorf tubes (7x total of ~1.5 mL, with remaining ~9.5 mL in foil-covered 50-mL conical tube
• Store at -20_o C
• Working concentration = 10-100 ug/mL