BCR 2.0

Cloning Strategy

Linker Optimization

Antibody Choices 

Immediate To Do's:

• look over syk linker in silico cloning 
• update timeline
• miniprep CD79A linkers miniprep
• TRE:mKate, pLac:mKate, hEF1a:LacI miniprep
• digests and gels 
• TRE:Gal4VP16 LR
• run SDM on gel -> gel extraction

BCR 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