Cloning Strategy

CD79A Linker Optimization

Antibody Choices 

Monday's To Do's 

•  Oligomerize beta-amyloid
•  Midi's:
  •  hEF1a:CymR
  •  hEF1a-CuO:mKate
  •  CMV5-CuO:mKate
  •  TRE:Syk
•  digest:
  •  hEF1a:CymR
  •  hEF1a-CuO:mKate
  •  CMV5-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

^^^Email: bcrpeeps@mit.edu

^^^Pass: grapefruit

Higher Level To Do's:

•  Decide on other antigen-variable regions we want to test  
•  Other experimental planning (on experimental page)
•  Parts list (excel sheet detailing specs/locations of parts)
  
•  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?