Preparing an Overnight Culture of E. coli

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

Preparing an Overnight Culture of E. coli

Preparing an Overnight Culture of E. coli

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

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

Preparing an Overnight Culture of E. coli