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='''LB agar plate preparing'''= ==Materials== *LB Miller agar powder (EMD chemical Inc.) *1 L Flask *Petri dish -- 20 plates / 500mL agar *Antibiotics - Kanamycin [http://en.wikipedia.org/wiki/Kanamycin](50mg / mL, 1000X, stored in -20 degree) ==Procedures== *Weigh the LB agar powder 37 g / 1 L -> 18.5 g / 500 mL. Dissolve 18.5 g LB agarose mix into 500 mL diH2O in a 1 L-flask. Makes two of these mixture. ** To stir the mixture for dissolving is not necessary. The autoclaving will help dissolving the mixture by itself. *Wrap the flask cap with foil and tape the autoclave-indicator tape. *Put the flasks on autoclave tray and autoclave the flask using the machine on 3rd floor. **Sign the log (name, date, which cycle been used, and machine condition) **Open the door by press "open". **Take out the upper iron tray if necessary. Put our mixture on the lower iron tray. Press 'close' to close the door. **Press '6' for selecting liquid cycle (121 degree, 30 min). Press 'start'. The total autoclaving time is about 45 min. **When the procedure is done. Press 'open' to open the gate. Wear gloves and take our media out. *Gently stirring the media for better mixing. Cool the Agar media in RT for 30 min or until it become 60 degree. At the same time thaw the antibiotics on ice. *Add 500 uL of antibiotics (1000X) into the flask (containing 500 mL agar media). Gently stirring the media for mixing. *Set up the gas-flaming. Pour the agar plate beside the flaming set. *Cool down the finished plates for 1 hr beside the flame. *Store the solid plates in sleeve in 4 degree. ----
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