Team:Macquarie Australia/WetLab/Protocols/Media

From 2014.igem.org

Revision as of 05:13, 16 October 2014 by Chharodn (Talk | contribs)

Protocols


Media & Plates

Well prepared media are crucial to successful selection and growth of organisms.

LB AGAR PLATE PREPARATION

  • Add 15g Bacto agar to 1000mL of LB media and autoclave (121oC for 15mins)
  • Add 1000uL of Chloroamphenicol (25mg/mL), Ampicillin (50mg/mL) or Kanamycin (30mg/mL) and mix well before plating out and setting the agar.

LB MEDIA

Ingredients: Tryptone 10g, Yeast extract 5g, NaCl 10g, Milli-Q water to 1000mL

Methods: Dissolved 10g tryptone, 5g yeast extract and 10g NaCl in 800mL Milli-Q water, making use of a magnetic stirrer. Once dissolved, bring volume up to 1 L using Milli-Q water. Autoclave 1L of the solution (121°C, 15 min, standard liquid cycle).

SOC MEDIA (FOR COMPETENT CELLS)

Ingredients: 10g bacto-tryptone, 2.5g bacto-yeast, 1000µl 5M NaCl, 417µL 3M KCl, 1.205g 20mM Mg2SO4, 805g 20mM D-glucose & 500mL Milli-Q water.

Methods: The adjusted quantities were combined in a 1 L measuring column with constant stirring and then placed in the autoclave for sterilisation (121°C, 15 min, standard liquid cycle).

Retrieved from "http://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Media"