https://eln.excellgen.com/index.php?action=history&feed=atom&title=Transfomation_for_making_glycerol_stock 2026-10-18T11:23:52Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=Transfomation_for_making_glycerol_stock&diff=20017&oldid=prev 2019-07-03T14:00:26Z

<p></p> <p><b>New page</b></p><div>== &lt;span dir=&quot;auto&quot;&gt;Creating Transfomation for making glycerol stock&lt;/span&gt; ==<br /> <br /> *take competent cells from minus 80 freezer<br /> *pipet 100 microliters and mix with 0.5 microliter of given plasmid<br /> *incubate on ice for 30 minutes<br /> *pour some water on the PCT PCR machine and heat up to setting &quot;42-30&quot; and put on it for about 30~45 seconds<br /> *take it out and let sit on ice for 2 minutes<br /> *add 750 microliters of SOC<br /> *put in 37 oC shaker incubator for about two hours<br /> *spin for 30 seconds<br /> *dump out liquid but leave about 50-100 microliters<br /> *then spray on LB/K plate and incubate<br /> *---------&gt; next day<br /> *wash out colonies with 1.5 ml&amp;nbsp;&amp;nbsp; &lt;s&gt;&lt;/s&gt;LB/K<br /> *spin down, remove ~0.75 ml liquid<br /> *add .75 mL of 80% glycerol<br /> *print label and enter into database<br /> *put into a freezer box in -80 freezer<br /> [[Category:Protocols]]</div>

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