Section Six - References & Budget
Author: Eric Schneider · 2026a-eric-schneider Node: Genspace NYC Affiliation: BioArt Studio, MakerSpace Charlotte
Form Prompts
List all references cited in your project documentation.
Provide a budget for your project. Include all costs associated with your project, including materials, equipment, and any other expenses. If you have not yet incurred any costs, provide an estimated budget.
References
All references ordered by first appearance across Sections 1–5. See Appendix A (separate file: biolight_appendixA_optogenetic_systems.md) for optogenetic systems evaluation table and associated references R17–R21.
Section 2 — Aims
R1. Li X, Zhang C, Xu X, Miao J, Yao J, Liu R, Zhao Y, Chen X & Yang Y. A single-component light sensor system allows highly tunable and direct activation of gene expression in bacterial cells. Nucleic Acids Research 48(6):e33 (2020). doi:10.1093/nar/gkaa044 · PMC7102963 · PMID 31989175 (eLightOn — BioLightV5 molecular ancestor. Primary citation.) Link to eLightOn
R2. Jin X & Riedel-Kruse IH. Biofilm Lithography enables high-resolution cell patterning via optogenetic adhesin expression. PNAS 115(14):3698–3703 (2018). doi:10.1073/pnas.1720676115 · RRID:Addgene_107741 (pDawn-sfGFP control construct — Addgene #107741.)
Section 3 — Background
R3. Kuldell N, Bernstein R, Ingram K, Hart KM. BioBuilder: Synthetic Biology in the Lab. O’Reilly Media (2015). ISBN 978-1491904299. https://www.oreilly.com/library/view/biobuilder/9781491904299/ (Karen Ingram co-author and scientific illustrator — introduced HTGAA 2026, leading to the BioLight project.)
R4. Levskaya A, Chevalier AA, Tabor JJ, Simpson ZB, Lavery LA, Levy M, Davidson EA, Scouras A, Ellington AD, Marcotte EM, Voigt CA. Synthetic biology: Engineering Escherichia coli to see light. Nature 438(7067):441–442 (2005). doi:10.1038/nature04405 · PMID 16306980 (Foundational bacterial photography — Cph8 chimeric receptor. Primary citation 1.)
R5. Tabor JJ, Salis HM, Simpson ZB, Chevalier AA, Levskaya A, Marcotte EM, Voigt CA, Ellington AD. A Synthetic Genetic Edge Detection Program. Cell 137(7):1272–1281 (2009). doi:10.1016/j.cell.2009.04.048 · PMID 19563759 · PMC2775486 (Tabor Lab edge detection — bacterial photography lineage.)
R6. Tabor JJ, Levskaya A, Voigt CA. Multichromatic Control of Gene Expression in Escherichia coli. J. Mol. Biol. 405(2):315–324 (2011). doi:10.1016/j.jmb.2010.10.038 (Tabor Lab CcaS/CcaR multichromatic — inline reference.)
R7. Teller E (Astro Teller, Captain of Moonshots, X / Google X). Quoted by Thomas L. Friedman in: Thank You for Being Late: An Optimist’s Guide to Thriving in the Age of Accelerations. Farrar, Straus and Giroux (2016). ISBN 978-0374273538. (“Today is the slowest rate of change we will ever experience.”)
R8. Gartner Research. Term “citizen data scientist” introduced ca. 2016. https://www.gartner.com/en/information-technology/glossary/citizen-data-scientist
R9. Mace RL. Universal Design: Barrier-Free Environments for Everyone. Designers West 33(1):147–152 (1985). Center for Universal Design, North Carolina State University College of Design. https://design.ncsu.edu/research/center-for-universal-design/ (Ron Mace coined Universal Design — mentor to Eric Schneider at NCSU.)
R10. The MIT License. Massachusetts Institute of Technology. https://opensource.org/licenses/MIT (Open-source license governing Photoplasm hardware, software, and protocol releases.)
Section 4 — Experimental Design
R11. Tabor JJ. Plate-based assays for light-regulated gene expression systems. Methods in Enzymology 497:373–391 (2011). doi:10.1016/B978-0-12-385075-1.00015-9 (Agarose slab embedding method — foundational protocol for bacteriography.)
R12. Sanders ER. Aseptic laboratory techniques: plating methods. Journal of Visualized Experiments (JoVE) 63:e3064 (2012). doi:10.3791/3064 (Bacterial plating methodology.)
R13. IDT Codon Optimization Tool. Integrated DNA Technologies. https://www.idtdna.com/CodonOpt (Used for RsLOV protein sequence → E. coli K12 codon-optimized DNA conversion during BioLightV5 design.)
R14. Addgene plasmid #107741 — pDawn-sfGFP. Deposited by Ingmar Riedel-Kruse Lab. RRID:Addgene_107741. https://www.addgene.org/107741/ (Control construct for Aim 2 — Test & Analyze. See also R2.)
Section 5 — Results & Validation
R15. Hurter F & Driffield VC. Photochemical investigations and a new method of determination of the sensitiveness of photographic plates. Journal of the Society of Chemical Industry 9:455–469 (1890). (H&D curve — foundational sensitometry. Conceptual framework for the bacterial H&D curve deliverable of Aim 2.)
R16. Li X, Zhang C, Xu X, Miao J, Yao J, Liu R, Zhao Y, Chen X & Yang Y. A single-component light sensor system allows highly tunable and direct activation of gene expression in bacterial cells. Nucleic Acids Research 48(6):e33 (2020). doi:10.1093/nar/gkaa044 (Full author list verified against published article. See also R1.)
Appendix A References
R17–R21 listed in full in biolight_appendixA_optogenetic_systems.md.
| Ref | Citation | Role |
|---|---|---|
| R17 | Olson EJ et al. Nature Methods 11(4):449 (2014) | CcaS/CcaR — evaluated, deselected |
| R18 | Jayaraman P et al. ACS Synth. Biol. 5(12):1363 (2016) | EL222 — evaluated, deselected |
| R19 | Baumschlager A & Khammash M. Advanced Biology 5(5):2000256 (2021) | Review — candidate selection |
| R20 | Multamäki E et al. ACS Synth. Biol. 11(10):3354 (2022) | pREDawn — evaluated, deselected |
| R21 | Castillo-Hair SM et al. Nature Commun. 10:3099 (2019) | Light-switchable systems — evaluated, deselected |
Reference Count Summary
| Section | New references introduced | Running total |
|---|---|---|
| 1 — Abstract | 0 | 0 |
| 2 — Aims | 2 (R1, R2) | 2 |
| 3 — Background | 8 (R3–R10) | 10 |
| 4 — Experimental Design | 4 (R11–R14) | 14 |
| 5 — Results & Validation | 2 (R15, R16) | 16 |
| 6 — Appendix A | 5 (R17–R21) | 21 |
Budget
Section 1 — BioLightV5 Plasmid & Genspace Lab Protocol
| # | Item | Description | Qty | Cost | Status |
|---|---|---|---|---|---|
| 1.01 | BioLightV5 clonal gene synthesis | Twist Biosciences — pBioLight-1B-eLightOn-v1, single clonal gene, pUC19 backbone, <5 kbp | 1 | $289 | ✅ Ordered |
| 1.02 | pDawn-sfGFP control construct | Addgene #107741 — bacterial stab, Riedel-Kruse Lab | 1 | $65 | ✅ Ordered |
| 1.03 | DH5α competent cells | NEB #C2987 or Genspace stock — high-efficiency transformation | 1 tube | $60 | ⏳ Pending — confirm at orientation |
| 1.04 | LB broth + LB agar | Standard bacteriological media for culture and plating | 1 L each | $20 | ⏳ Pending — confirm Genspace stock |
| 1.05 | Ampicillin | Selection antibiotic for AmpR BioLightV5 and pDawn | 1 g | $15 | ⏳ Pending — confirm Genspace stock |
| 1.06 | SOC medium | Recovery medium post-transformation | 50 mL | $10 | ⏳ Pending — confirm Genspace stock |
| 1.07 | Low-melt agarose | NuSieve GTG or equivalent — agarose slab embedding for bacteriography | 5 g | $25 | ⏳ Pending — confirm Genspace stock |
| 1.08 | Petri dishes 90 mm | Standard BSL-1 plates for transformation and exposure | 20 | $10 | ⏳ Pending — confirm Genspace stock |
| 1.09 | Miniprep kit | Plasmid preparation from overnight culture | 1 kit | $60 | ⏳ Pending |
| 1.10 | Sanger sequencing | Verification of BioLightV5 construct — 2–3 reactions | 3 | $30 | ⏳ Pending — method TBD at orientation |
| 1.11 | Glycerol | Working stock banking at −80°C | 100 mL | $8 | ⏳ Pending — confirm Genspace stock |
| 1.12 | Sterile tubes, tips, consumables | General BSL-1 bench consumables | lot | $20 | ⏳ Pending — confirm Genspace stock |
| 1.13 | Genspace community lab membership | Community lab member rate — prorated or one month of activity (TBD at orientation May 28) | 1 month | $TBD | ⏳ Pending — confirm May 28 |
Section 1 Subtotal: ~$612 + TBD (membership)
Section 2 — Photoplasm Device Hardware
| # | Item | Description | Qty | Cost | Status |
|---|---|---|---|---|---|
| Optical Platform | |||||
| 2.01 | Besseler darkroom enlarger | Vintage used enlarger — column, baseboard, negative carrier, lamphouse, dual condenser lens array, focusing lens all included. Repurposed as Photoplasm bio-imaging optical platform | 1 | $20 | ✅ Purchased |
| Compute & Control | |||||
| 2.02 | Raspberry Pi 5 (4GB) | Main compute and control unit — Raspberry Pi OS Bookworm 64-bit | 1 | $60 | ✅ Purchased |
| 2.03 | MicroSD card 64GB | OS and project storage | 1 | $12 | ✅ Purchased |
| 2.04 | USB-C power supply 5V/5A | Pi 5 power — separate from 12V LED rail | 1 | $15 | ✅ Purchased |
| LED Light Ring | |||||
| 2.05 | Cree XP-E2 470nm LED stars | 3-Up star, 470nm blue, from LEDsupply.com | 3 | $11 | ✅ Purchased |
| 2.06 | Carclo diffuser optics | 26.5mm frosted wide-spot polycarbonate lens, LEDsupply.com | 3 | $10 | ✅ Purchased |
| 2.07 | Aluminum heat sink | Two-piece housing for 3-Up LED + optic, LEDsupply.com | 3 | $18 | ✅ Purchased |
| 2.08 | Cooling fan kit | Active thermal management for LED array at rated drive current | 1 | $15 | ✅ Purchased |
| 2.09 | 6.8Ω 2W resistors | Current limiting at 12V rated drive for Cree XP-E2 | 3 | $8 | ⏳ Ordered |
| 2.10 | 120Ω resistors | String current limiting — 3 strings of 3 series LEDs | 3 | $3 | ✅ Purchased |
| 2.11 | 12V DC power supply | LED rail power — separate from Pi 5V supply | 1 | $18 | ✅ Purchased |
| 2.12 | EBOOT LED ring | Consumer-grade 470nm ring — calibration reference, superseded by Cree | 1 | $12 | ✅ Purchased |
| PWM / MOSFET Driver | |||||
| 2.13 | IRLZ44N MOSFET | N-channel logic-level MOSFET — PWM driver for LED ring | 5 | $8 | ✅ Purchased |
| 2.14 | 10kΩ resistors | Gate pull-down resistors | 10 | $3 | ✅ Purchased |
| 2.15 | 470Ω resistors | Gate protection resistors | 10 | $3 | ✅ Purchased |
| 2.16 | 330Ω resistors | GPIO protection resistors | 10 | $3 | ✅ Purchased |
| 2.17 | Breadboard half-size | Working prototype platform | 2 | $8 | ✅ Purchased |
| 2.18 | Protoboard / perfboard | Permanent circuit construction from breadboard design | 2 | $10 | ⏳ Pending |
| 2.19 | JST PH 2.0mm connectors + crimp tool | Standardized connector system across all modules | 1 kit | $22 | ✅ Purchased |
| Digital Image Mask | |||||
| 2.20 | OLED SSD1309 128×64 SPI | Current digital image mask — Waveshare breakout, mounted in negative carrier position | 1 | $18 | ✅ Purchased |
| 2.21 | ILI9341 transmissive LCD | Planned upgrade — true non-emissive variable density mask | 1 | $25 | ⏳ Planned — post Aim 2 |
| Optical Accessories | |||||
| 2.22 | 515nm long-pass filter | sfGFP emission separation from 470nm excitation | 1 | $28 | ✅ Purchased |
| Spectral Sensor | |||||
| 2.23 | AS7341 11-channel spectral sensor | Waveshare breakout — I2C, 256X gain, F2+F3 470nm dose proxy | 1 | $25 | ✅ Purchased |
| Incubation Control | |||||
| 2.24 | PTCYIDU PTC heater element | Plate heater — 37°C setpoint, variable and tunable | 1 | $18 | ✅ Purchased |
| 2.25 | DS18B20 temperature probe | 1-Wire temperature sensor — GPIO4 | 1 | $8 | ✅ Purchased |
| 2.26 | IRLZ44N MOSFET (heater) | PWM heater control — GPIO13 | 2 | $4 | ✅ Purchased |
| Camera | |||||
| 2.27 | Raspberry Pi Camera Module 3 | Machine vision feedback during duty cycle — Aim 2 installation | 1 | $25 | ⏳ Ordered |
| Enclosure & Fabrication | |||||
| 2.28 | PETG filament gray bulk — Bambu | Gray PETG, bulk rolls — all Photoplasm enclosure parts | 4 rolls | $56 | ✅ Purchased |
| 2.29 | Bambu X1 Carbon print time | Fabrication at MakerSpace Charlotte | ~8 h | $0 | ✅ MakerSpace membership |
| 2.30 | M2.5 hardware (screws, nuts) | Assembly hardware for all enclosure components | 1 lot | $8 | ✅ Purchased |
| 2.31 | Jumper wires, ribbon cable | GPIO wiring between all components | 1 lot | $12 | ✅ Purchased |
| 2.32 | LED matrix board PCB | biolight_board2_led_matrix_alpha_01 — 120×120×2mm PETG, 9×9 diagonal RGB | 1 | $15 | ⏳ In progress |
Section 2 Subtotal: ~$528
Total Project Budget Summary
| Section | Description | Subtotal | Status |
|---|---|---|---|
| Section 1 | BioLightV5 Plasmid & Genspace Lab Protocol | ~$612 + TBD | ⏳ Partially ordered |
| Section 2 | Photoplasm Device Hardware | ~$528 | ⏳ Mostly purchased |
| Project Total | ~$1,140 + TBD | ⏳ In progress |
TBD = Genspace community lab membership — prorated community member rate, to be confirmed at orientation May 28, 2026. Several Genspace consumable line items (1.03–1.12) may be available from Genspace lab stock at reduced or no cost — to be confirmed at orientation. Separate optical component sourcing alternatives are deferred to Aim 3 — Learn & Refine (Visionary) as community-driven hardware innovation. All estimates subject to revision as actuals are received.
Status key:
- ✅ Purchased — actual cost incurred
- ✅ Ordered — committed, actual confirmed
- ⏳ Ordered — committed, awaiting delivery
- ⏳ Pending — not yet ordered, confirmation needed
- ⏳ Planned — scheduled for future aim phase
- ⏳ In progress — design underway, cost estimated