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 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)



Sequencing account information:
^^^Email: bcrpeeps@mit.edu

^^^Pass: grapefruit

 

 

Experimental Progress:

Cloning Progress
Parts Tree
Cloning Strategy
Experiments
Linker Optimization
Antibody Choices 

 

Useful Links: 

Protocols
Immediate To Do's:

BCR 2.0 To Do

^^^Email: bcrpeeps@mit.edu

^^^Pass: grapefruit

Expand
titleLinker optimization
TaskClaimed byDateOther Notes
design CD79B SDM primersLA CRJun 16 
  • order SDM primers
CRJun 16 
  • SDM + transform
 Jun 18 
  • pick colonies
 Jun 19 
  • miniprep
 Jun 20 
  • digest
 Jun 20 
  • sequencing
 Jun 19 
LR LacI BCR Jun 16 
  • kill LR and transform
 Jun 17 
  • pick colonies
 Jun 18 
  • miniprep
 Jun 19 
  • digest
 Jun 19 
  • sequencing
 Jun 19 
Expand
titleVariable region construction
TaskClaimed byDate CompletedOther Notes
Variable region cloning planning   
    

## 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  
  •  Other experimental planning (on experimental page)
  •  How do we do site directed mutagenesis?
  •  Literature research: 
    •  kozak sequences for heavy and light chains and CD79s
    •  truncated TEVp
    •  other antigens to detect using BCR (we were thinking of small molecule detection)
    •  inducible promoters (we were considering LacI - an inducible repressor - but we want to look into other options first) 
  •  Cloning planning 
    •  hEF1a:Syk-pTet:mRFP-TEVp
    •  variable region 
  • Is gantenerumab proprietary and can we actually publish with it?