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Team:Macquarie Australia/WetLab/Protocols/Media
From 2014.igem.org
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<h3>Media & Plates</h3> | <h3>Media & Plates</h3> | ||
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| - | <h4>LB AGAR PLATE PREPARATION</h4> | + | <p><i>Well prepared media are crucial to successful selection and growth of organisms.</i></p> |
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| - | <ul style="list-style-type: decimal;"> | + | <h4>LB AGAR PLATE PREPARATION</h4> |
| - | <li>Add 15g Bacto agar to 1000mL of LB media and autoclave ( | + | |
| - | <li>Add 1000uL of Chloroamphenicol (25mg/mL), Ampicillin (50mg/mL) or Kanamycin (30mg/mL) and mix well before plating out and setting the agar. </li> | + | <ul style="list-style-type: decimal;"> |
| - | </ul | + | <li>Add 15g Bacto agar to 1000mL of LB media and autoclave (121<sup>o</sup>C for 15mins).</li> |
| - | + | <li>Add 1000uL of Chloroamphenicol (25mg/mL), Ampicillin (50mg/mL) or Kanamycin (30mg/mL) and mix well before plating out and setting the agar. </li> | |
| + | </ul> | ||
| - | <h4>LB MEDIA</h4> | + | <h4>LB MEDIA</h4> |
| - | <p> | + | <p><b>Ingredients:</b> Tryptone 10g, Yeast extract 5g, NaCl 10g, Milli-Q water to 1000mL</p> |
| - | <b>Ingredients:</b> Tryptone 10g, Yeast extract 5g, NaCl 10g, Milli-Q water to 1000mL | + | <p><b>Methods:</b> 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<sup>o</sup>C, 15 min, standard liquid cycle). </p> |
| - | </p> | + | |
| - | <p> | + | |
| - | <b>Methods:</b> 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 ( | + | |
| - | </p> | + | |
| - | <h4>SOC MEDIA (FOR COMPETENT CELLS)</h4> | + | <h4>SOC MEDIA (FOR COMPETENT CELLS)</h4> |
| - | <p> | + | <p><b>Ingredients:</b> 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.</p> |
| - | <b>Ingredients:</b> 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. | + | <p><b>Methods:</b> The adjusted quantities were combined in a 1 L measuring column with constant stirring and then placed in the autoclave for sterilisation (121<sup>o</sup>C, 15 min, standard liquid cycle).</p> |
| - | </p> | + | |
| - | <p> | + | |
| - | <b>Methods:</b> The adjusted quantities were combined in a 1 L measuring column with constant stirring and then placed in the autoclave for sterilisation ( | + | |
| - | </p> | + | |
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</div> | </div> | ||
</section> | </section> | ||
</body> | </body> | ||
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Revision as of 05:26, 16 October 2014
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 (121oC, 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 (121oC, 15 min, standard liquid cycle).
