Troubleshooting: E. coli Overnight Culture Setup 2014

Preparing an Overnight Culture of E. coli 2014

Possible Problems • When there is a mixed culture of red and white (nonexpressing) cells, the white cells will grow faster than those that are using their resources producing red fluorescent protein (mFP) = less mFP produced for purification. • Some transformed cells stop expressing rpf fairly early.

Materials • Sterile 125 mL flasks of LB/amp broth. • Vented cap to ensure adequate aeration during overnight shaking • Mini shaker table • Frozen cells transformed with pARA-R • Tube of sterile arabinose (500mg/mL)

Procedure Morning, Day before Lab 6 • Aseptically, add 500 uL of transformed cells into each flask of LB/amp broth • Secure vented cap (do not wet the membrane) • Shake and incubate (35o C) for 2–3 hours • Following the 2–3 hours, add the appropriate volume of arabinose to each flask so the final concentration of arabinose is 5mg/100mL • Continue to shake overnight

Time vs Growth

Troubleshooting: E. coli Overnight Culture Setup 2014

Troubleshooting: E. coli Overnight Culture Setup 2014

Presentation Transcript

Stool Culture, E. coli O157:H7

E. Coli bacteria

Stool Culture, E. coli O157:H7

E. Coli Testing

The Outbreak of E. Coli

Chiral Expansion of E. coli Summary

E. coli

Return of the E. coli

E. coli Genome

Cells of E. coli bacteria

Cling- E. coli

Preparing an Overnight Culture of E. coli

T4 bacteriophage infecting an E. coli cell

Checking Out The Culture Of An Overnight Summer Camp

Checking Out The Culture Of An Overnight Summer Camp

e coli o121

E. Coli

e coli advantages