Editing Transfomation for protein expression

= <span dir="auto">Creating Transfomation for protein expression</span> =
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*take competent cells (Nico21(DE3)pLysN) from minus 80 freezer
*pipet 100 microliters and mix with 0.5 microliter of given plasmid
*incubate on ice for 30 minutes
*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
*take it out and put on ice, let sit on ice for 2 minutes
*add 750 microliters of SOC
*put in shaker for about two hours
*spin for one minute
*dump out liquid but leave about 50-100 microliters
*suspend bacteria with pipette
*spray on LB/KC plate and spread with glass beads
*put in incubator overnight, agar side up
*---------> next day
*wash out colonies with LB/KC or LB/AC and pipette into a 50 mL plastic tube
*add LB/KC or LB/AC until it reaches 15 mL
*put in shaker for about 2-3 hours
*add to 1 or 2 Liters of LB/KC or LB/AC
*grow in shaker
*induce with 0.5 mL of 1 M IPTG per bottle
*induce at 37 oC for 4 hours, or room temperature overnight
*spin down cells, combine, weigh cell pellet weight and store cells at -80 freezer
[[Category:Protocols]]