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Team:Macquarie Australia/WetLab/Protocols/Media
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<li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/PlasmidPreps">Plasmid Prep</a></li> | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/PlasmidPreps">Plasmid Prep</a></li> | ||
<li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/SDSPAGE">SDS-PAGE</a></li> | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/SDSPAGE">SDS-PAGE</a></li> | ||
| + | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Heat">Heat Shock</a></li> | ||
| + | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Operons">Induction of Operons</a></li> | ||
</ul> | </ul> | ||
</div> | </div> | ||
<div class="cont-out"> | <div class="cont-out"> | ||
| + | |||
<h3>Media & Plates</h3> | <h3>Media & Plates</h3> | ||
| - | <p></p> | + | |
| + | <p><i>Well prepared media are crucial to successful selection and growth of organisms.</i></p> | ||
| + | |||
| + | <h4>LB AGAR PLATE PREPARATION</h4> | ||
| + | |||
| + | <ul style="list-style-type: decimal;"> | ||
| + | <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> | ||
| + | |||
| + | <img src="https://static.igem.org/mediawiki/2014/3/35/Media.png" height=300 /><img src="https://static.igem.org/mediawiki/2014/d/d7/Media2.png" height=300 /> | ||
| + | <p><b>Figure 1.</b> Preparation of LB agar plates.</p> | ||
| + | |||
| + | <h4>LB MEDIA</h4> | ||
| + | <ul> | ||
| + | <li><b>Ingredients:</b> Tryptone 10g, Yeast extract 5g, NaCl 10g, Milli-Q water to 1000mL.</li> | ||
| + | <li><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). </li> | ||
| + | </ul> | ||
| + | |||
| + | <h4>SOC MEDIA (FOR COMPETENT CELLS)</h4> | ||
| + | <ul> | ||
| + | <li><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. </li> | ||
| + | <li><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).</li> | ||
| + | </ul> | ||
| + | |||
</div> | </div> | ||
</section> | </section> | ||
</body> | </body> | ||
</html> | </html> | ||
Latest revision as of 13:09, 17 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.
Figure 1. Preparation of LB agar plates.
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).
