Difference between revisions of "Team:NYU Shanghai/Protocols"
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<p><em>Note:</em> Some formulations of SOB use 10 mM MgCl2 and 10 mM MgSO4 instead of 20 mM MgSO4. | <p><em>Note:</em> Some formulations of SOB use 10 mM MgCl2 and 10 mM MgSO4 instead of 20 mM MgSO4. | ||
SOB medium is also available dry premixed from Difco, 0443-17.<br>Adjust to pH 7.5 prior to use. This requires approximately 25 ml of 1M NaOH per liter.<br><br><b>15/10 medium</b><br><br>Growth of competent TOP10 cells in Example 2 of the Bloom05 patent is performed in 15/10 broth, which is similar to SOB:</p> | SOB medium is also available dry premixed from Difco, 0443-17.<br>Adjust to pH 7.5 prior to use. This requires approximately 25 ml of 1M NaOH per liter.<br><br><b>15/10 medium</b><br><br>Growth of competent TOP10 cells in Example 2 of the Bloom05 patent is performed in 15/10 broth, which is similar to SOB:</p> | ||
− | |||
<li>1.5% yeast extract</li> | <li>1.5% yeast extract</li> | ||
<li>1% Bacto-Tryptone</li> | <li>1% Bacto-Tryptone</li> | ||
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<li>10 mM MgCl2</li> | <li>10 mM MgCl2</li> | ||
<li>10 mM MgSO4</li> | <li>10 mM MgSO4</li> | ||
− | |||
<br> | <br> | ||
<table border="1"> | <table border="1"> | ||
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<li>37ºC incubator</li> | <li>37ºC incubator</li> | ||
<li>Shaker</li> | <li>Shaker</li> | ||
+ | </p> | ||
+ | <p> | ||
<br><br>Procedure | <br><br>Procedure | ||
<ol> | <ol> | ||
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<li>You can pick a single colony, make a glycerol stock, grow up a cell culture and miniprep.</li> | <li>You can pick a single colony, make a glycerol stock, grow up a cell culture and miniprep.</li> | ||
</ol> | </ol> | ||
+ | </p> | ||
+ | <p> | ||
<br>Controls | <br>Controls | ||
<li>Competent cells on LB plate</li> | <li>Competent cells on LB plate</li> | ||
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<div id="mpText" style="display:none"> | <div id="mpText" style="display:none"> | ||
<p> | <p> | ||
− | Estimated time | + | Estimated time: 40 minutes |
<br><br>Materials | <br><br>Materials | ||
<li>Biomiga Miniprep Kit</li> | <li>Biomiga Miniprep Kit</li> |
Revision as of 14:44, 17 September 2015