Team:Georgia State/Collaborations
Backyard Biology @ Emory University: Project JBroth
Low Cost LB and Competent Cells!
Our interest in our topic began when we were researching possible projects. What we found most interesting was a research paper from the Journal of Pakistan Medical Association that tested the efficacy of microwaving LB rather than autoclaving. Also, a project that inspired us was the project from the University of Calgary (2014) that tested the efficacy of a homemade wash for competent cells. From these projects, we asked ourselves: Could one conduct a synthetic biology project without access to a professional laboratory?
This summer, we created a cost effective alternative to lab grade Luria Broth (LB). To compare our homemade broth, J-Broth, and lab grade LB we compared optical densities of liquid cultures and transformation efficiency of competent cells. For each method of testing, we compared autoclaved LB, microwaved LB, and microwaved J-Broth.
Materials/Methods
We grew XL1-Blue E. coli cells in liquid culture of lab grade luria broth and J-broth. The media was either autoclaved or microwaved for 8 minutes at 650 Watts before the cells were added.
We then made competent cells using XL1-Blue e. coli cells that were grown in each type of media following the Inoue method (Inoue, 1990) and calculated the transformation efficiency for comparison.
Results
Price Comparison
Lab grade LB Powder (1L) LB Powder (Sigma-Aldrich) : $141.50 for 1000 grams $0.1415/gram x 25 = $3.538Total = $3.538
Lab Grade LB Individual Ingredients(1L)
Tryptone (Sigma-Aldrich) : $615 for 1000 grams $0.615/gram x 10 = $6.15Yeast Extract : $190 for 1000 grams $0.19/gram x 5 = $0.95
Sodium Chloride (Sigma-Aldrich) : $71.50 for 1000 grams $0.0715/gram x 10 = $0.715
Total = $7.815
J-broth(1L)
Bulk Supplements Amino Acids - $16.96 for 250 grams $0.067/gram x 10 =$0.67Marmite Yeast Extract - $12.63 for 500 grams $0.025/gram x 5 = $0.125
Sodium Chloride (table salt) - $8.28 for 1000grams $0.0083/gram x 10 = $0.083
Total = $0.275
Savings from LB Powder = $3.263/liter
Savings from LB Ingredients = $7.54/liter
Put tables here!!!
LB Type | OD 600 Diluted by 10 |
Lab Autoclave | 0.122 |
Lab Microwave | 0.185 |
J-Broth Microwave | 0.098 |
LB Type | Transformation Efficiency (CFU/microgram) |
Lab Autoclave | 1.2e6 |
Lab Microwave | 3.16e6 |
J-Broth Microwave | 1.17e6 |