Cloning Strategy

CD79A Linker Optimization

Antibody Choices 

STATUS TABLE

 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

• 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 

•  Cytometry time 
•  Order Cumate 
•  Chicken lysozyme, gp120
•  Antibodies for antigen binding detection

Waiting on primers to do list:

• 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?

THINGS TO NOT FORGET TO DO:

• SYK-15tTEVP golden gate when we figure out what's wrong with the AarI
• 3T3 MIDI