Biomolecular Breadboards:Protocols
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{| border = "1" cellpadding="5" style="width: 800px" | {| border = "1" cellpadding="5" style="width: 800px" | ||
! scope="col" | Simple circuit | ! scope="col" | Simple circuit | ||
| - | ! scope="col" | Assembly time for ''in vitro'' testing | + | ! scope="col" | Assembly time for ''in vitro'' testing (linear DNA) |
| + | ! scope="col" | Assembly time for ''in vitro'' testing (multiple plasmids) | ||
| + | ! scope="col" | Assembly time for ''in vitro'' testing (single plasmid) | ||
! scope="col" | Assembly time for ''in vivo'' testing | ! scope="col" | Assembly time for ''in vivo'' testing | ||
(1 plasmid construct) | (1 plasmid construct) | ||
|- | |- | ||
| - | | style="width: 200px; | Negatively autoregulated gene | + | | style="width: 200px; | Negatively autoregulated gene |
| + | | style="text-align:center" | 6.5 hours | ||
| + | | style="text-align:center" | N/A | ||
| + | | style="text-align:center" | 3 days | ||
| + | | style="text-align:center" | 3 days | ||
|- | |- | ||
| - | | Bistable switch || style="width: 200px; text-align:center" | 3 days || style="width: 200px; text-align:center" | 5 days | + | | Bistable switch |
| + | | style="width: 200px; text-align:center" | 6.5 hours | ||
| + | | style="width: 200px; text-align:center" | 3 days | ||
| + | | style="width: 200px; text-align:center" | 5+ days* | ||
| + | | style="width: 200px; text-align:center" | 5+ days* | ||
|} | |} | ||
Revision as of 11:56, 20 November 2012
| Home | Protocols | DNA parts | Preliminary Data | Models | More Info |
Cell-free circuit breadboard
The following protocols are available for the cell-free circuit breadboard:
- Reaction Preparation - Cell-free expression of circuits from linear or plasmid DNA
- TXTL template, v2.3 - TXTL spreadsheet for designing experiments
Cost estimate
The cell-free circuit breadboard costs approximately 2.64 cents per uL, or 31.7 cents per 12 uL reaction. The cost distribution is shown in the table below.
| Component | Material price per
μL of reaction | Labor price per
μL of reaction | Total price per
μL of reaction | Total price per
12 μL reaction |
|---|---|---|---|---|
| E. coli crude extract | 0.45 cents | 0.62 cents | 1.07 cents | 12.84 cents |
| Amino acids | 0.21 cents | 0.03 cents | 0.24 cents | 2.93 cents |
| 3-PGA | 0.39 cents | 0.94 cents | 1.33 cents | 15.96 cents |
| Total price | 1.05 cents | 1.6 cents | 2.64 cents | 31.7 cents |
For a more detailed cost estimate, please see Breadboard Cost Estimate.
Design Cycle Time Analysis
| Simple circuit | Assembly time for in vitro testing (linear DNA) | Assembly time for in vitro testing (multiple plasmids) | Assembly time for in vitro testing (single plasmid) | Assembly time for in vivo testing
(1 plasmid construct) |
|---|---|---|---|---|
| Negatively autoregulated gene | 6.5 hours | N/A | 3 days | 3 days |
| Bistable switch | 6.5 hours | 3 days | 5+ days* | 5+ days* |
( * Estimated. All other times are measured. )
For a more detailed description, please see Breadboard Cycle Time Measurements.
