Difference between revisions of "Team:NYU Shanghai/Protocols"
Line 305: | Line 305: | ||
<p> | <p> | ||
<ol> | <ol> | ||
− | <li>Calculate total amount of volume of LB agar you would need to use | + | <li>Calculate total amount of volume of LB agar you would need to use. You need about 25mL for every 10cm plate and 10mL for every 6cm plate.</li> |
− | + | ||
− | + | ||
</ul> | </ul> | ||
</li> | </li> | ||
<li>For every 100mL of LB agar needed, measure out 4g of LB Agar Powder. | <li>For every 100mL of LB agar needed, measure out 4g of LB Agar Powder. | ||
− | |||
− | |||
− | |||
</li> | </li> | ||
− | <li>Put the agar powder in a flask | + | <li>Put the agar powder in a flask. Use a much bigger flask to prevent overflow when autoclaiving. </li> |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</li> | </li> | ||
− | <li>Put deionized water into the flask up to the desired volume. | + | <li>Put deionized water into the flask up to the desired volume. <strong>Do not mix.</strong> LB Agar is not supposed to dissolve in room temperature. If you mix the powder it will get stuck on the walls of the flask</li> |
− | + | ||
− | + | ||
− | + | ||
</li> | </li> | ||
<li>Autoclave. </li> | <li>Autoclave. </li> | ||
− | <li>After autoclave, | + | <li>After autoclave, move to the hood. Let the LB agar cool until it’s less than 60ºC or until you can touch the flask. While waiting, you can start labeling the plates. <strong>DO NOT LET THE AGAR SOLIDIFY</strong></li> |
<li>Insert any antibiotics/sugars needed. If you are not sure about the ratio, you can follow this simple example: | <li>Insert any antibiotics/sugars needed. If you are not sure about the ratio, you can follow this simple example: | ||
<p style="text-align: center;"><b>50mL LB + 500µL Arabinose* + 50µL Ampicillin**</b><span style="font-size: 10;"><br><i> | <p style="text-align: center;"><b>50mL LB + 500µL Arabinose* + 50µL Ampicillin**</b><span style="font-size: 10;"><br><i> |
Revision as of 14:41, 17 September 2015