Making MEM Media

Procedure

Take 25 ml pipet and peel back wrapping and connect to pipet aid. Put the wrapping paper in the black trash bag. Remove cap off FBS tube and draw up 25 ml of serum in 25 ml pipet.

Remove cap off 500 ml Media bottle and add 25 ml of FBS serum into the media. Repeat the last two steps to add 25 ml more of FBS serum into the media. Toss FBS tubes and 25 ml pipet into the orange biohazard bag.

Now take 10 ml pipet and connect it to the pipet aid barrel. Carefully remove the wrapping paper and put it into the black trash bag. Make sure the tip of pipet is facing away from you and not touching any non-sterile surface.

Without sliding hands over tube, remove cap off 15 ml tube and draw up 10 ml of PSG and mix by pipetting up and down to mix the precipitate. After precipitate dissolves, draw up 5 ml of PSG and and add it to the MEM media bottle.

Put the cap back on PSG tube and label it with your name and return it to the freezer for future use. Toss 10 ml pipet into the orange biohazard bag.

Label the MEM media bottle with initials and 10% FBS+PSG. Place the media bottle in refrigerator for future use or can be used immediately.

Clean the hood area with 70% ethanol and turn it off.Turn off the water bath and make sure everything is clean.

Result

Conclusion

• The first trial of this lab was not successful because of experimental errors. Media was not properly made because more than required amount of Penn/Strep/Glutamine was added accidently.

• The second trial of this lab was successful because correct amount of materials were added to MEM media and media was properly made.