Cloning Strategy

CD79A Linker Optimization

Antibody Choices 

STATUS TABLE

 To do's:

•  Midiprep (from freezer)
  •  S9TEV
  •  B15T15
•  Golden Gate
  •  B9T9
  •  B15T9
•  LR

Monday's To Do's 

•  Oligomerize beta-amyloid
•  Midi's:
  •  hEF1a:CymR
  •  hEF1a-CuO:mKateGmab H
  •  CMV5hEF1a-CuO:mKate
  •  TRE:Syk
   digest:
  • Gmab L
  •  hEF1a-CuO:CymRCD79A
  •  hEF1a-CuO:mKateCD79B
  •  CMV5hEF1a-CuO:mKate 
  •  TRE:Syk
•  Make Sodium Azide solution 
•  Pick colonies:
  •  TRE:Syk-15-tTEVp golden gate
  •  Golden gate donor mini product
  •  LRs:
    •  EGSH:mKate
    •  hEf1a:VgEcR-2A-RXR
    •  TRE:mKate
    •  hEF1a:Gal4VP16 (remember which pDEST)
    •  hEF1a:CD79B-TCS-Gal4VP16 
    •  hEF1a:tagBFP

Non-Lab To Do's:

•  Order chicken lysozyme and gp120?
•  order antibodies for antigen binding detection

Geneious To Do List:

•  Antibody cloning for chicken lysozyme - Kathryn 
•  hef1a sequencing primersBCR 2.0 To Do

• • 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?