====== Making Liquid Media ======

 --- //[[celiser12@gmail.com|Christine Robbins]] 2018/11/14 14:00//

==== Description ====
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Below are general directions for making liquid media. Please review them whenever making liquid stocks of media. 

**Notes:**
  * Whenever using media, please remember that if you open the last bottle, you need to make more right away. Not later. If you are a student and you must leave for class, please let your friendly neighborhood Lab Tech or Lab Aide know that the media needs to be made
  * It is always recommended that you print out the media recipe and cross off every item as you add it to avoid mistakes  
  * Carbon sources should always be added after autoclaving as the media is being used


====Directions====

  * Grab the appropriately sized, sterilized [[beakers|beaker]] and [[bottles|bottles]] from the [[glassware|glassware shelves]]
        * For liquid media, a beaker the same size as your final volume is sufficient
        * You will only fill bottles to the last labeled volume, for example: A 1L bottle is only labeled to 900mL, therefore you will only fill it to 900mL
  * Label all glassware with the name of the media you are making, your initials and the date (yes, this includes the beaker)
  * Place a stir bar in the bottom of the beaker 
        * The stir bar should be approximately 1/3 the diameter of the beaker
        * When removing the stir bar from the autoclave pouch, remove it by pushing it out, not by sticking your fingers in and grabbing it. 
  * Add [[milliq_water|MilliQ Water]] to approximately 1/4 the final volume using a [[graduated_cylinders|graduated cylinder]] and turn on the magnet on the hot plate
  * Weigh out the reagents listed on the recipe and add them to the media one at a time 
        * It is best to add all reagents slowly to avoid clumping  
  * When all ingredients have been added, fill the beaker to the final volume with MilliQ Water using a graduated cylinder
  * When all ingredients are incorporated, pour the media into the labeled bottles
        * You will only fill bottles to the last labeled volume, for example: A 1L bottle is only labeled to 900mL, therefore you will only fill it to 900mL
        * Please pour carefully and avoid pouring out the stir bar
  * Cap the bottles so that the caps still wiggle, but do not come off (allows steam to escape without having exploding bottles
  * Place a piece of autoclave tape over the top of the cap and down the side of the bottle as seen in the image 
  * Autoclave the bottles on the liquids cycle
  * While the autoclave is running, clean up 
        * Clean up the chemical bench and scales with water followed by 70% EtOH
        * Throw away any used weigh boats
        * Wash the beaker and lab scoops 
  * When the autoclave is done, remove the bottles
  * When the bottles have cooled, screw the cap on completely and place them on the shelf where they belong