Brett 3-10-10

To see how other Plates are made follow this link

Plate pouring protocol (for 1000 ml)

Add a Teflon stir bar to a 2 liter or larger Erlenmeyer flask

Add 200 mL of dH2O from the milliQ ("Aqua") carboy followed by 20 grams peptone, 10 grams yeast extract, and 20 grams agar. See part numbers below.

Add 750 mL additional water to the flask and stir with no heat for a few minutes

Autoclave on the last liquid cycle (cycle 6) in the autoclave facility . You can either give it to Justin and ask him to bring it up when it comes out, or if he's not around set it yourself and set a timer to go down and get it.

After removing from autoclave, place flask on heat block and stir at low/medium heat then ADD GLUCOSE to 2%.  Our stock is usually 50% glucose .

Add any other component to the media or for plates of varying concentrations add component to 25 ml of molten YPD in a 50 ml conical tube, or 100ml in a small beaker or flask. For many compounds you will need to let the YEPD cool as much as possible before adding. When you can leave the back of your hand against the YEPD flask without wanting to poull kit away, it has cooled enough typically- be sure of this for your compound of interest though.  Label plates before or immediately after pouring.

Dump molten media into sterile Petri dishes and cover immediately- about 25ml per normal (10cm) plate.

Let dry overnight, then stack, bag, and place in cold room or on media shelf as needed for the type of media.

Part Numbers :

Peptone: Becton-Dickinson 211677

Yeast Extract: Becton-Dickinson 212750

Bacto Agar: Becton-Dickinson 214010

Glucose: Sigma 59378 or Bio-World

Our empty plates (petri dishes) are sterile 100X15mm BD Falcon 351029 plates.


YEPD Ingredients