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Today's To Do's:
  • restreak TRE:Syk-mRFP-(t)TEVp - eric 
  • miniprep digest verify TRE:Syk-mRFP-(t)TEVp
  • transform pENTR LacI and pENTR pLac
  • pick hEF1a:LacI, TRE:mKate, pLac:mKate
  • digest, gel hEF1a:LacI, TRE:mKate, pLac:mKate LR2
  • miniprep TRE:gal4 transformation 2 
  • digest, gel verify TRE gal4 transformation 2
  • split cells - Lyla 
  • pLac:mKate pick LR colonies - Lyla 
  • Inducible promoter LRs
  • ScaI to lab stock
Wednesday's To Do's
  • pick colonies: 
    • pLac:mKate 
  • Transform: 
    • Inducible promoter LRs
    • pENTR LacI and pENTR pLac
  • Miniprep, digest, gel 
    • TRE:Syk-pTet:mRFP-TEVp
Geneious To Do List:
  • SDM primers: CD79B, Gal4VP16, TEVp

  • TRE:gal4vp16 sequencing primers
  • TRE:Syk-mRFP-(t)TEVp sequening primers - 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 

 

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