Making MEM Media

Lab 1: Making Media for Cells

In this lab, the preparation of the sterile MEM media was conducted. This media will be used throughout the semester to maximize the growth of the HEK 293 cells.

Materials

500ml of MEM media

(Minimum Essential Media)

2x 25ml of Fetal Bovine Serum

5ml of Penicillin, Streptomycin and Glutamine mixture

Pipetman Aid

Level II Biosafety Cabinet

A few 10ml and 25ml pipets

37°C water bath

Pre-Lab Preparation

Biosafety Cabinet Set Up

Prior to beginning the lab, the bio safety cabinet was sterilized by turning on the blower and the UV lights. The UV illumination was on for 15 minutes, after which the working lights and outlets were turned on.

Water Bath Set Up

The water bath was also turned on in order to warm up the water to 37°C. This will thaw all of the components of the MEM media.

Sterile Technique

Throughout this lab and all labs moving forward the sterile technique will be applied. This technique minimizes the possibility of contamination, by washing the biosafety cabinet with ethanol as well as the containers of the materials that will be used in the experiment. The equipment such as pipets will also be open in a sterile method.

Making the MEM Media

Starting with the first 25ml of FBS, it was titrated to resuspend any possible sedimentation. It was then pipetted into the 500ml of media. This was then repeated with the second falcon tube of 25ml FBS. The 25ml pipet was then discarded into the biohazard container. With the use of a 5ml pipet the PSG mixture was titrated to resuspend it. 5ml were then drawn up and transferred into the 500ml MEM container.

MEM Media Storage

After completing the addition of all the components, the MEM media container was labeled 10% FBS +PSG. It was then stored in 4°C refrigerator, in order to preserve the media.

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