BCR 2.0
Switchable Antigen-Sensing System (S.A.S.S.)
Friday's To Do's
- Midi, digest, gel
- TRE:Syk-6-tTEVp
- Mini, digest
- TRE:Syk-15-tTEVp
- Mini, sequence
- B-3T3-G4
- B-3T9-G4
- B-6T6-G4
- B-6T9-G4
- B-9T3-G4
- B-9T6-G4
- B-9T12-G4
- B-12T6-G4
- B-12T9-G4
- B-12T12-G4
- B-15T3-G4
- B-15T15-G4
- Resend for sequencing
- A-3T15-G4
- A-9T15-G4
- TRE:Syk-9-tTEVp
- TRE:Syk-12-tTEVp
- Golden Gate, transform
- hEF1a:CD79A-15T9-G4 (because even though we midi'ed it, sequencing was bad)
- TRE:Syk-linker-TEVp (3, 6, 9, 12)
- hEF1a:CD79B-3T15-G4
- hEF1a:CD79B-9T9-G4
- Streak onto new plate (some white colonies hidden with red ones)
- hEF1a:CD79B-6T12-G4
- hEF1a:CD79B-15T9-G4
- Pick colonies
- pENTR HyHEL10 Heavy
- hEF1a:CD79B-9T15-G4
Thursday's To Do's
- Mini, Midi, digest, gel
- TRE:Syk-15-tTEVp mini #1,#2
- TRE:Syk-3-tTEVp midi
- A-15T9 midi
- Sequencing
- HyHEL10 H #2
- TRE:Syk-pTET:mRFP-TEVp (Syk3 read)
- Pick CD79B colonies
- Inoculate midi Syk-6-tTEVp
Wednesday's To Do's
- Golden gates, ligate, transform:
- CD79B-linker-G4
- Midi
- A-12T12
- Mini, digest, gel
- TRE:S-3-tT - don't digest this, we have a good one
- TRE:S-6-tT
- TRE:S-9-tT
- TRE:S-12-tT
- Pick
- TRE:Syk-15-tTEVp
- Innoculate midi:
- A15T9
- T:S3tT
- Analyze sequencing
- CD79B-RFP good
- A-3T15 bad
- A-9T15 bad
- A-15T9 good
- T:S-3-tTp (throw out new cultures + duplicate of mini)
- Sequenicng
- A-3T15 #2
- A-9T15 #2
- S-9-tT #1
- S-12-tT #1
Sequencing account information:
^^^Email: bcrpeeps@mit.edu
^^^Pass: grapefruit
Non-Lab To Do's
- Cytometry time
- Order Cumate
- Chicken lysozyme, gp120
- Antibodies for antigen binding detection
Waiting on primers to do list:
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?
THINGS TO NOT FORGET TO DO:
- SYK-15tTEVP golden gate when we figure out what's wrong with the AarI
- 3T3 MIDI