Biomolecular Breadboards:Protocols: Difference between revisions
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** [[Biomolecular Breadboards:Protocols:3-PGA Buffer Prep|3-PGA Buffer Preparation, 14X]] | ** [[Biomolecular Breadboards:Protocols:3-PGA Buffer Prep|3-PGA Buffer Preparation, 14X]] | ||
* [[Media: TXTL_v2_3.xlsx | TXTL template, v2.3]] - TXTL spreadsheet for designing experiments | * [[Media: TXTL_v2_3.xlsx | TXTL template, v2.3]] - TXTL spreadsheet for designing experiments | ||
We are the in the process of publishing a paper in the ''Journal of Visualized Experiments'' ([http://www.jove.com JoVE]) that describes this protocol in much more detail. The JoVE paper will be an open access paper. If you would like a preprint, contact [[Biomolecular Breadboards:More Info|Richard Murray]]. | |||
=== Cost estimate === | === Cost estimate === | ||
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{| border = "1" cellpadding="5" style="width: 800px" | {| border = "1" cellpadding="5" style="width: 800px" | ||
|- | |- | ||
| | ! rowspan=2 width=250px | Circuit | ||
! colspan=4 | Assembly time | |||
|- | |||
! ''In vitro'' testing <br>(linear DNA) | |||
! ''In vitro'' testing <br>(multiple plasmids) | |||
! ''In vitro'' testing <br>(single plasmid) | |||
! ''In vivo'' testing <br>(single plasmid) | |||
|- | |||
| Negatively autoregulated gene | |||
| style="text-align:center" | 6.5 hours | | style="text-align:center" | 6.5 hours | ||
| style="text-align:center" | N/A | | style="text-align:center" | N/A | ||
| style="text-align:center" | 3 days | | style="text-align:center" | 3 days | ||
| style="text-align:center" | 3 days | | style="text-align:center" | 3 days* | ||
|- | |- | ||
| Bistable switch | | Bistable switch | ||
| style="width: 200px; text-align:center" | 6.5 hours | | style="width: 200px; text-align:center" | 6.5 hours* | ||
| style="width: 200px; text-align:center" | 3 days | | style="width: 200px; text-align:center" | 3 days | ||
| style="width: 200px; text-align:center" | 5 | | style="width: 200px; text-align:center" | 5 days* | ||
| style="width: 200px; text-align:center" | 5+ days* | | style="width: 200px; text-align:center" | 5+ days* | ||
|} | |} | ||
<nowiki>* Estimated time based on similar constructions. All other times are measured.</nowiki> | |||
For a more detailed description, please see [[Biomolecular Breadboards:Protocols:Cycle Time Measurements|Breadboard Cycle Time Measurements]]. | For a more detailed description, please see [[Biomolecular Breadboards:Protocols:Cycle Time Measurements|Breadboard Cycle Time Measurements]]. | ||
Revision as of 10:04, 11 May 2013
| 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
We are the in the process of publishing a paper in the Journal of Visualized Experiments (JoVE) that describes this protocol in much more detail. The JoVE paper will be an open access paper. If you would like a preprint, contact Richard Murray.
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
| Circuit | Assembly time | |||
|---|---|---|---|---|
| In vitro testing (linear DNA) |
In vitro testing (multiple plasmids) |
In vitro testing (single plasmid) |
In vivo testing (single plasmid) | |
| Negatively autoregulated gene | 6.5 hours | N/A | 3 days | 3 days* |
| Bistable switch | 6.5 hours* | 3 days | 5 days* | 5+ days* |
* Estimated time based on similar constructions. All other times are measured.
For a more detailed description, please see Breadboard Cycle Time Measurements.
