Cloning Strategy

CD79A Linker Optimization

Antibody Choices 

STATUS TABLE

 To do's

:

•  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)

•  Transform golden gates - pick colonies later in the day 
•  
  
•   

Wednesday's to Dos

•  Redigest basically everything from yesterday and gel
•  Pick colonies
  •  TRE:Syk-15-tTEV
  •  hEF1a:CD79B-RFP-G4
  •  pENTR VRC01 heavy
  •  pENTR VRC01 light
  •  pENTR HyHEL-10 heavy
  •  pENTR HyHEL-10 light
•  Fix - see here for ideas
  •  TRE:Syk-15-tTEV
  •  hEF1a:CD79B-RFP-G4
  •  Syk linkers
•  Miniprep andn digest
  •  CD79A-3T15
  •  CD79A-12T9
  •  CD79A-15T9
•  Inoculate midipreps if the sequencing comes back good
•  Regolden gate things - extra ligation for BsaI golden gate

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