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)

Monday's To Do's 

•  Oligomerize AB
•  Miniprep, digest, gel
  •   CD79A linker constructs
  •  Midis
•  transform IgM-RFP golden gate
•  golden gate TRE:Syk-15-TEVp in gg donr

Tuesday's To Do's

•  Gel for oligo annealing
  •  Order new annealing buffer
•  Pick RFP-IgM Colony
•  Syk-15-TEV GG

Saturday's To Do's 

•  Midipreps 

•  CD79A golden gate T4 ligation

•  Transformations
  •  CD79A golden gates 
  •  IgM RFP golden gate

•  3T3

•  6T6

•  6T12

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:

•  TRE:Syk-15-TEVp
•  CD79B-mRFP-G4 PCR

• 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?