PGLO Lab Jan. 2013 | Page 8

Methodology

Sample preparation

make 500 mL of lb agar (~40 plates)

To Begin...

8

Weigh out the following into a 1L Erlenmeyer flask:

5g NaCl

5g Tyrptone

2.5g Yeast Extract

7.5g Agar

Add dH2O to reach 500 mL

Place Erlenmeyer flask into a pressure cooker...

Allow pressure to build until the solution reaches 20 PSI, for about

20 minutes. This will cause the bacteria to die and the solution will

become sterile.

Pour the solution into the petri dishes!

*(See page 9 for details on how to distribute the solution)*

Leave the petri dishes out at room temperature for 1 day and...

Allow the solution in the petri dish to dry out, or "cure", so the

moisture will leave. The solution within the petri dish will now be

ready to absorb any added substances.

Place petri dishes into a refrigerator until they are ready for use!

REHYDRATE ARABINOSE AND AMPICILLIN

In order to rehydrate the sugar and antibiotic, 3mL of transformation solution must be added directly into each vial, using a sterile pipette.