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
Beckman Coulter
EMD Millipore
MP Biomedicals
Roche Applied Science
Sigma Aldrich
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