Wet Lab Pages
From GcatWiki
This is the space for MWSU and DC wet lab students to create content. Our part numbers will be in this range only: BBa_K091000 to BBa_K091999.
- Davidson Wet Lab Protocols
- MWSU Wet Lab Protocols
- Build a LuxR Repressor PDF Reprint
- Plasmid Compatability
- Davidson -80 Stocks
- Using Amp and Amp^R as communication
- FAQs for Wikis
Registry parts to clone
Things to Do
- Experiment with making lines of media in petri dishes.
- Determine concentration of HSL needed to turn on our receiver + reporter. Compare with BBa_T9002
- Determine what percent of a colony need to be sender cells in order for an entire colony to become switched on by HSL.
- We need to make sender and receiver cells (using existing sub-parts such as F1610), and/or take from registry (e.g., BBa_T9002)
- Design experiments to test the distance HSL and beta lactamase can diffuse in agar and over what time spans.
- Make a derived pLac promoter that will permit LuxR+HSL to be a repressor to be used in combination with Lux pR promoter. See this paper.Andrew Gordon, Robert Cool
- Make an AmpR sender device. Look for AmpR basic part in Registry.
Composite Parts:
| Part | Description | Student Responsible (DC or MW) | Location and Vector | Resistance | Results from 5/22/08 | miniprep |
|---|---|---|---|---|---|---|
| BBa_F2621 | AHL receiver with lux pR and codes for LuxR | Karlesha Roland (DC) | 1001 12B pSB1A2 | amp | growth MW | no |
| BBa_I15030 | Lux-sender(Autoinducing)Codes for LuxI and LuxR | Kelly Davis (DC) will this send to part BBa_I13263 | 1015 8F pSB3K3 | kan | growth MW | no |
| BBa_I13263 | Lux Receiver (HSL & R0063 driven)produces YFP | Pallavi Penumetcha (DC) | 1002 8E pSB1A2 | amp | growth MW & DC | yes 210ng/ml |
| BBa_f2622 | 3OC6HSL receiver that outputs PoPS | James Barron (DC) | 1001 12E pSB1A2 | amp | no growth | no |
| BBa_F1610 | Device that receives PoPS and outputs LuxI | Malcolm Campbell (DC) | 1016 10D pSB1AK3 | amp & kan | growth MW | no |
| BBa_I0426 | Las-reciver(EYFP) | Kristi Muscalino (DC) | 1008 5B pSB1A2 | amp | growth MW & DC | yes 210ng/ml |
| BBa_I0407 | LasI test | Erin Feeney (DC) | 1008 4F pSB1A2 | amp | growth MW & DC | yes 76.37ng/ml |
| BBa_I0466 | RhlR Protein Generator without LVA | Laurie Heyer (DC) | 1001 5C pSB1A2 | amp | growth MW | yes 129.7ng/ml |
Basic Parts:
| Part | Description | Student Responsible (DC or MW) | Location and Vector | Resistance | Results from 5/22/08 | miniprep |
|---|---|---|---|---|---|---|
| BBa_C0061 | LuxI gene with LVA | Madeline Parra (DC) | 1000 3E pSB1A2 | amp | growth MW | |
| BBa_C0161 | LuxI gene without LVA | Xiao Zhu (MW) | 1001 8B pSB1A2 | amp | ||
| BBa_C0062 | LuxR gene without LVA | John Igo (MW) | 1000 3F pSB1A2 | amp | ||
| BBa_R0063 | lux pL | Aaron Lewis (MW) | 1000 4H pSB1A2 | amp | ||
| BBa_R0062 | lux pR | Andrew Gordon(MW) | 1000 4G pSB1A2 | amp | ||
| BBa_C0070 | rhlI gene | Jeff Poet (MW) | 1013 3E pSB2K3 | kan | ||
| BBa_C0071 | rhlR gene with LVA | Max Win (DC) | 1013 3F pSB2K3 | kan | no growth | |
| BBa_C0171 | rhlR gene without LVA | MWSU | 1001 6F pSB1A2 | amp | ||
| BBa_R0079 | las pR | Todd Eckdahl (MW) | 1001 10F pSB1A2 | amp | ||
| BBa_C0079 | lasR gene | Robert Cool (MW) | 1013 4F pSB2K3 | kan | ||
| BBa_C0078 | LasI gene | Alicia Allen (MW) | 1013 4E pSB2K3 | kan | ||
| BBa_J07019 | fecA promoter with Fur box | Not in Registry | ||||
| BBa_#### | Description | Sven(MW) | ||||
| BBa_#### | Description | Sven(MW) | ||||
| BBa_#### | Description | Sven(MW) |
Fluorescent Protein Gradient Testers:
| Part | Description | Student Responsible (DC or MW) | Location and Vector | Resistance | Results | miniprep |
|---|---|---|---|---|---|---|
| BBa_J04450 | Lac-RBS-RFP-T | Robert Cool (MW) | 1004 4f pSB1A2 | amp | ||
| BBa_J04451 | pLac-RBS-RFP w/ LVA tag-T-T | Erin Feeney and Kelly Davis (DC) | 1016 9F pSB1AK3 | amp and kan | ||
| BBa_I13520 | pBAD-RBS-mRFP-T-T | Erin Feeney and Kelly Davis (DC) | 1009 7B pSB1A2 | amp | ||
| BBa_J5528 | pBAD-RBS-GFP-T-T | Erin Feeney and Kelly Davis (DC) | 1015 7A pSB2K3 | kan | ||
| BBa_J04430 | pLac-RBS-GFP-T-T | Erin Feeney and Kelly Davis (DC) | 1004 4H pSB1A2 | amp |
Parts for Sending and Receiving Signals:
| Part | Description | Student Responsible (DC or MW) | Location and Vector | Resistance | Results | miniprep |
|---|---|---|---|---|---|---|
| BBa_J07019 | fec A promoter with fur box | James Barron (DC) | Not in Registry-Needs to be amplified | |||
| BBa_F2622 | 3OC6HSL Receiver Device | Pallavi Penumetcha (DC) | 1001 12e pSB1A2 | amp | ||
| BBa_I14032 | pLac promoter | Pallavi Penumetcha (DC) | 1013 8e pSB2K3 | kan | ||
| BBa_S03632 | lacI promoter and luxI gene | Pallavi Penumetcha (DC) | 1016 7f pSB1AK3 | amp and kan | ||
| BBa_E0240 | RBS and GFP | Pallavi Penumetcha (DC) | 1001 4B pSB1A2 | amp |
