BCR 2.0
Switchable Antigen-Sensing System (S.A.S.S.)
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Friday
To do's
To Do's- CD79B-RFP-G4 SDM
- Midiprep and digest
- hEF1a:tagBFP
- Inoculate minis
- RFP-IgM CH golden gate
- hEF1a:CD79A-linker-G4
- Miniprep and digest
- hEF1a:CD79A-linker-G4
- TRE:Syk-15-tTEV
- hEF1a:VP16Gal4
- EGSH:mKate
- hEF1a:VgEcR-2A-RxR
- hEF1a:CD79B-TCS-G4
- TRE:mKate
- Inoculate midis
- hEF1a:VP16Gal4
- EGSH:mKate
- hEF1a:VgEcR-2A-RxR
- hEF1a:CD79B-TCS-G4
- TRE:mKate
- Sequencing
- hEF1a:CD79A-linker-G4
- TRE:Syk-15-tTEV
- TRE:Syk-RFP-TEV (missing read only)
- TRE:Syk-RFP-tTEV (missing read only)
Thursday's To Do's
- Innoculate midi cultures
- hEF1a:tagBFP (from liquid mini culture)
- Inoculate mini cultures - because picking from plates of transformed reactions
- hEF1a:CD79A-linker-G4 golden gates
- Pick some for growth at 37C and others for growth at 30C
- TRE:Syk-15-tTEV - pick a couple of these (it's a golden gate with no screening)
- TRE:mKate
- hEF1a:gal4VP16
- EGSH:mKate
- hEF1a:VgEcR-2A-RxR
- hEF1a:CD79B-TCS-Gal4vp16
- Transform
- RFP-IgM CH golden gate
- Replate failed CD79A linker golden gates
- Ordering things
- Chicken lysozyme, gp120
- Antibodies for antigen binding detection
- Cumate?
Geneious To Do List:
- Antibody cloning for chicken lysozyme - Kathryn
hef1a sequencing primersBCR 2.0 To Do
:
- Midiprep (from freezer)
- S9TEV
- B15T15
- Golden Gate
- B9T9
- B15T9
- LR
- hEF1a-CuO:Gmab H
- hEF1a-CuO:Gmab L
- hEF1a-CuO:CD79A
- hEF1a-CuO:CD79B
- hEF1a-CuO:Lyn
- Digest all of the Syk-TEVp and Syk-tTEVp midis just to be sure (since there's an internal cut site that didn't get read in sequencing)
Sequencing account information:
^^^Email: bcrpeeps@mit.edu
^^^Pass: grapefruit
## Digest tTEVp and TEVp midis just to be sure (since there's an internal cut site)
Non-Lab To Do's
- Bug Brian about ordering antibody testing supplies
Higher Level To Do's:
- Decide on other antigen-variable regions we want to test
- Is gantenerumab proprietary and can we actually publish with it?