BCR 2.0

Cloning Strategy

CD79A Linker Optimization

Antibody Choices 

Monday's To Do's 

• pick more CD79A linker colonies (all old DNA is not good)
• pick CD79A SDM colony 
• make more DNA of failed constructs from 07/03 gel
• digest, gel:
  •  pENTR mKate
  • hEF1a:CD79B-TCS-G4
  • pENTR LacI
  • hEF1a:Gal4VP16
• LR EGSH:mKate, hEf1a:VgEcR-2A-RXR
• sequence:
  • TRE:syk-ptet:mrfp-(t)TEVP
  • hEF1a:CD79B-TCS-G4
• transform:
  • MAV1212 because we don't have enough 
  • TRE:syk because we don't have a midi

CMV5-CuO 715:

hEF1a-CuO 715:09

CymR-CuO 710:09

Geneious To Do List:

• Antibody cloning for chicken lysozyme - Kathryn 
• hef1a sequencing primers

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