3 - Bacterial growth
3 - Bacterial growth kdorfman Fri, 08/07/2020 - 15:44Grow bacteria
Streak frozen cells on agar plate
Pick a colony and grow overnight in liquid LB (plus antibiotic if necessary)
Concentrate ON culture in centrifuge
Make 2 dilution series: 10-fold, 2-fold
2 fold
- tubes 1A - 10A
- 0.5 mL LB in each
- add .5 mL culture to first, mix thoroughly
- serially dilute by moving 0.5 mL along the series*
10 fold
- tubes 1B - 10B
- 0.9 mL LB in each
- add .1 mL culture to first, mix thoroughly
- serially dilute by moving 0.1 mL along the series
Take OD measurements
- Load 250 uL of each dilution into 96 well plate
- samples in 1st 10 wells of 1st 2 rows
- LB in last 2 wells
- Read, using bug OD
Plot results
- Put fraction of original culture next to each OD
- Cut, paste special, values only
- Sort data
- Make a scatterplot
- Find the lowest concentration just above the noise for hemocytometer
Hemocytometer
- Load 10 uL into hemocytometer.
- Photograph.
- Also take photographs of too concentrated and too dilute samples
Colony Counts
- spread 100 uL from last 5 10-fold dilutions on agar plates
- Grow overnight
- Photograph