Dulbeccos Modified Eagle's Media, DMEM adding extras

Dulbeccos Modified Eagle's Media, DMEM adding extras

• It is best to prepare a few tubes with the required amount of FETAL BOVINE SERUM (FBS) and Penicilin Streptomycin (Pen-Strep), and store them in the freezer. Each time only unfreeze the small tubes, instead of the whole bottle. As for 500 ml DMEM, it requires 50 mL (or 10%) of FBS and 5 mL (or 1%) of Penicilin Streptomycin (Pen-Strep).
• Step 1: Before prepare for DMEM solution, take out 1 tube of FBS and 1 tube pen-strep from the freezer and place them in the fridge (1 day in advance, after the overnight is fine).
• Step 2: Work inside the LAF bench and use syringes and 0.2 µm filter to add 50 ml FBS and 5 ml Pen-Strep to the 500ml DMEM.
• Tight the caps, and slightly shake the bottle to mix them.  Label the bottle with names and date, then store it in the fridge.

Stock solution preparation:

FETAL BOVINE SERUM (FBS)  

• Step 1:  Use 75% antibac ethanol alcohol to spray both hands with gloves, then the surface of the FBS bottle, the surface of 25 ml Serological Pipette, a few of 50 ml conical tube, and place them inside the LAF fume hood.  
• Step 2: Inside the LAF hood, completely loose the conical tube caps and placed them in frame holder. Transfer 51 ml FBS into each conical tube, and tight the caps.  
• Step3: label each conical tube, put them in a plastic bag, and store them in the freezer. 

  Penicilin Streptomycin (Pen-Strep)  

• Use the same procedure as FBS, and each conical tube with 5,5 ml Pen-strep, then store them in the freezer with labels.