Today we are making e.coli for our DDW experiments. The first step is to make the e.coli medium. We made up a small batch of LB Broth, using the instructions from the Sigma-Aldrich website. We only made 100mL of LB Broth, instead of 1L, as follows:
- Mixed 2 g of LB Broth powder with 100mL of distilled water.
- Autoclave for 15 minutes at 121°C.
After the mixture cooled, we measured 10mL into two test tubes and mixed in a minute amount of e.coli.
Next we placed the test tubes in the New Brunswick Scientific Innova 4300 Incubator Shaker at 37° C. Our shaker’s thermostat is about .3°C – .4°C high , so we stuck a thermometer probe in the shaker to get an accurate temperature measurement.
We set the incubator shaker at 70 RPM for a few hours. I have to leave for class, and Anthony wasn’t sure if we needed to leave it longer or maybe even overnight. Please check his blog for the final details.