Biomolecular Breadboards:Protocols:cost estimate
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Cell-Free Circuit Breadboard Cost Estimate
The cell-free circuit breadboard costs approximately 1.05 cents per uL for materials, or 12.6 cents per typical 12 uL reaction. Including the labor cost of preparing the reaction components, it costs approximately 2.64 cents per uL, or 31.7 cents per 12 uL reaction. The material cost distribution among the reaction components is shown in the table below. The percent volumes listed were used to calculate the approximate price of each component per reaction uL. The labor cost distribution can be found at start-up and labor costs.
Reaction Ingredients
| Component | % Reaction Volume | Price per uL of reaction | Price per 12 uL reaction | Source | Catalog Number |
|---|---|---|---|---|---|
| Extract | 33.3% | $0.00448 | $0.05376 | Homemade | |
| Amino acids | 25% | $0.0021 | $0.0252 | Homemade | |
| 3-PGA buffer | 7.14% | $0.00391 | $0.04697 | Homemade | |
| Mg-glutamate | 1% | negligible | Sigma-Aldrich | 49605 | |
| K-glutamate | 1.3% | negligible | Sigma-Aldrich | G1149 | |
| DTT | 1% | negligible | Sigma-Aldrich | D0632 | |
| PEG8000 | 5% | negligible | Promega | V3011 | |
| DNA, water, inducer | User-supplied | ||||
| Total Cost | $0.0105 | $0.126 |
E. coli crude extract, amino acids, and 3-PGA buffer are "homemade" components, in that they cannot be bought directly from a vendor but must be prepared ahead of time by the user. The cost breakdown for each is detailed below.
E. coli crude extract
"Amount per 3X prep" refers to the amount of crude extract made following the protocol E. Coli Crude Extract Preparation, 3X. On average, one prep yields 10.5 mL of extract, which is enough for about 31500 uLs of cell-free circuit reaction.
| Ingredient | Bulk Quantity | Bulk Price | Amount per 3X prep | Price per 3X prep | Source | Catalog Number |
|---|---|---|---|---|---|---|
| 2xYT | 1 kg | $87.3 | 139.93 g | $12.216 | MP biomedicals | 3012-032 |
| Bacto-Agar | 1 lb | $171.11 | 0.44 g | $0.17 | BD (from Fisher) | 214010 |
| Chloramphenicol powder | 5 g | $30.50 | 3.876 mg | $0.02 | Sigma-Aldrich | C1919 |
| DTT | 5 g | $97.40 | 0.927 g | $18.06 | Sigma-Aldrich | D0632 |
| Ethanol (100%) | 500 mL | $54.68 | 114 uL | $0.01 | Fisher | BP2818 |
| K-glutamate | 500 g | $161.50 | 48.8 g | $15.76 | Sigma-Aldrich | G1149 |
| Mg-glutamate | 250 g | $47.10 | 21.76 g | $4.10 | Sigma-Aldrich | 49605 |
| Potassium phosphate monobasic solution | 1 L | $43.80 | 90.15 mL | $3.95 | Sigma-Aldrich | P8709 |
| Potassium phosphate dibasic solution | 1 L | $38.00 | 163.92 mL | $6.23 | Sigma-Aldrich | P8584 |
| Tris | 1 kg | $133.58 | 13.325 g | $1.78 | Fisher | BP1521 |
| Total Price | $62.30 |
| Disposable Material | Bulk Quantity | Bulk Price | Amount per 3X prep | Price per 3X prep | Source | Catalog Number |
|---|---|---|---|---|---|---|
| Beads (0.1 mm diameter) | 1 lb | $34.00 | 80 g | $6.00 | BioSpec | 11079101 |
| Bead beating tubes | 500 count | $141.00 | 42 count | $11.84 | BioSpec | 522S |
| Bead filters | 100 count | $112.00 | 6 count | $6.72 | Bio-Rad | 732-6204 |
| Bio-rad Bradford protein assay kit (BSA) | 1 kit | $123.00 | 1 assay | $0.62 | Bio-Rad | 500-0201 |
| Cuvettes | 500 count | $170.30 | 7 count | $2.38 | Fisher | 14-955-127 |
| Dialysis cassettes | 10 count | $85.40 | 6 count | $51.24 | Thermo Scientific (from Fisher) | 66380 |
| Total Price | $78.80 |
| Total Price of E. coli crude extract, 3x Prep (Ingredients + Disposable Materials) | $141.10 |
| Price per uL of reaction | $0.00448 |
| Price per 12 uL reaction | $0.05376 |
Amino acids
One kit yields about 41.6 mL of amino acids, which is enough for approximately 166,400 uLs of cell-free circuit reaction. See Amino Acid Preparation for instructions on how to prepare the kit.
| Ingredient | Quantity | Price | Price per uL of reaction | Price per 12 uL reaction | Source | Catalog Number |
|---|---|---|---|---|---|---|
| RTS Amino Acid Sampler | 1 kit | $349.00 | $0.0021 | $0.0252 | 5 Prime | 2401530 |
3-PGA buffer
"Amount per 1-fold prep" refers to the amount of 3-PGA buffer made following the instructions for 1-fold preparation of 3-PGA buffer in the protocol 3-PGA Buffer Preparation, 14X. 1-fold preparation yields 427 uL of 3-PGA buffer, which is enough for 5977 uLs of cell-free circuit reaction.
| Ingredient | Bulk Quantity | Bulk Price | Amount per
1-fold prep |
Price per
1-fold prep |
Source | Catalog Number |
|---|---|---|---|---|---|---|
| HEPES KOH ph8 | 100 g | $156.50 | 71.493 mg | $0.11 | Sigma Aldrich | H6147 |
| ATP | 1 g | $51.50 | 5.6 mg | $0.29 | Sigma Aldrich | A8937 |
| GTP | 1 g | $767.00 | 5.155 mg | $3.95 | USB (from Affymetrix) | 16800 |
| UTP | 1 g | $177.00 | 3.19 mg | $0.56 | USB (from Affymetrix) | 23160 |
| CTP | 1 g | $298.00 | 3.0727 mg | $0.92 | USB (from Affymetrix) | 14121 |
| tRNA (E. coli) | 100 mg | $148.00 | 1.2 mg | $1.78 | Roche Applied Science | MRE600 |
| CoA | 25 mg | $81.80 | 1.2 mg | $3.93 | Sigma Aldrich | C4282 |
| NAD | 250 mg | $37.60 | 1.43 mg | $0.22 | Sigma Aldrich | N6522 |
| cAMP | 1 g | $185.50 | 1.76 mg | $0.33 | Sigma Aldrich | A9501 |
| Folinic Acid | 100 mg | $64.60 | 0.208 mg | $0.13 | Sigma Aldrich | F7878 |
| Spermidine | 1 g | $50.80 | 0.000871 g | $0.04 | Sigma Aldrich | 85558 |
| 3-PGA | 1 g | $230.50 | 0.0476 g | $10.97 | Sigma Aldrich | P8877 |
| Tris | 1 kg | $133.58 | 1.302 g | $0.17 | Fisher | BP1521 |
| Total Price of 3-PGA buffer, 1-fold prep | $23.40 |
| Price per uL of reaction | $0.00391 |
| Price per 12 uL reaction | $0.04697 |
Start-up and labor costs
The cost of the cell-free reaction is calculated assuming the user works in a typical biology laboratory. It does not include the cost of common laboratory supplies and equipment that the average lab could reasonably be expected to have. However, preparation of the E. coli extract will probably require an initial investment in a few uncommon items. They are listed below, and comprise the "start-up cost" of the cell-free circuit breadboard.
| Item | Quantity Needed | Total Price | Vendor | Catalog Number |
|---|---|---|---|---|
| Mini-Beadbeater | 1 | $885 | BioSpec | 3110BX |
| BL21 Rosetta 2 (E. coli strain) | < 0.4 mL | $98.00 | Novagen (from EMD Millipore) | 71402 |
| 4L Erlenmyer flasks | 6 | $590.52 | Fisher | 26500-4000 |
| Total Start-Up Cost | $1,573.52 |
Components of the cell-free reaction require time to prepare, and cannot be simply ordered from a vendor like other in vitro expression kits. To fairly compare reaction costs, we provide below a rough labor cost estimate. It assumes an employee is paid $14/hr, and is working alone. The time listed is the approximate time needed to prepare a 3X prep of E. coli crude extract, 1 kit of amino acids, and a 1-fold prep of 3-PGA buffer.
| Component | Time per prep | Cost per prep | Cost per uL | Cost per 12 uL |
|---|---|---|---|---|
| E. coli crude extract | 14 hrs | $196 | $0.0062 | $0.075 |
| Amino acids | 4 hrs | $56 | $0.00034 | $0.004 |
| 3-PGA | 4 hrs | $56 | $0.0094 | $0.112 |
| Total Cost | $0.016 | $0.191 |
Vendor Links
| 5 Prime | |
| Affymetrix | http://www.affymetrix.com |
| Beckman Coulter | https://www.beckmancoulter.com |
| Bio-Rad | http://www.bio-rad.com/ |
| BioSpec | http://www.biospec.com/ |
| EMD Millipore | http://www.emdmillipore.com/ |
| Fisher | http://www.fishersci.com/ |
| MP Biomedicals | http://www.mpbio.com/ |
| Promega | http://www.promega.com/ |
| Roche Applied Science | http://www.roche-applied-science.com/ |
| Sigma Aldrich | http://www.sigmaaldrich.com/ |
