Transfomation for making glycerol stock

Creating Transfomation for making glycerol stock

• take competent cells 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 seconds
• take it out and let sit on ice for 2 minutes
• add 750 microliters of SOC
• put in 37 oC shaker incubator for about two hours
• spin for 30 seconds
• dump out liquid but leave about 50-100 microliters
• then spray on LB/K plate and incubate
• ---------> next day
• wash out colonies with 1.5 ml   LB/K
• spin down, remove ~0.75 ml liquid
• add .75 mL of 80% glycerol
• print label and enter into database
• put into a freezer box in -80 freezer