https://eln.excellgen.com/index.php?action=history&feed=atom&title=Transfomation_for_protein_expressionTransfomation for protein expression - Revision history2026-10-18T09:33:02ZRevision history for this page on the wikiMediaWiki 1.44.0https://eln.excellgen.com/index.php?title=Transfomation_for_protein_expression&diff=20019&oldid=prevCrm at 14:05, 3 July 20192019-07-03T14:05:37Z<p></p>
<p><b>New page</b></p><div>= <span dir="auto">Creating Transfomation for protein expression</span> =<br />
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*take competent cells (Nico21(DE3)pLysN) 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 "42-30" and put on it for about 30~45&nbsp; seconds<br />
*take it out and put on ice, let sit on ice for 2 minutes<br />
*add 750 microliters of SOC<br />
*put in shaker for about two hours<br />
*spin for one minute<br />
*dump out liquid but leave about 50-100 microliters<br />
*suspend bacteria with pipette<br />
*spray on LB/KC plate and spread with glass beads<br />
*put in incubator overnight, agar side up<br />
*---------> next day<br />
*wash out colonies with LB/KC or LB/AC and pipette into a 50 mL plastic tube<br />
*add LB/KC or LB/AC until it reaches 15 mL<br />
*put in shaker for about 2-3 hours<br />
*add to 1 or 2 Liters of LB/KC or LB/AC<br />
*grow in shaker<br />
*induce with 0.5 mL of 1 M IPTG per bottle<br />
*induce at 37 oC for 4 hours, or room temperature overnight<br />
*spin down cells, combine, weigh cell pellet weight and store cells at -80 freezer<br />
[[Category:Protocols]]</div>Crm