Cloning Strategy

CD79A Linker Optimization

Antibody Choices 

Tuesday's To Do'

•  Oligomerize AB

•  Miniprep, digest, gel
  •   CD79A linker constructs
  •  Midis

•  transform IgM-RFP golden gate

s

•  Gel for oligo annealing
  •  Order new annealing buffer
•  CD79B, gal4 PCR
•  Golden Gates:
  •  hEF1a:CD79B-pTET:mRFP-Gal4 golden gate
  •  TRE:Syk-15-
  TEVp in gg donr

Tuesday's To Do's

•  Gel for oligo annealing
  •  Order new annealing buffer
•  Pick
  • tTEVp
•  TRE:Syk-pTET:mRFP-tTEVp sequencing 
•  Pick pENTR RFP-IgM Colony
•  Syk-15-TEV GG

Saturday's To Do's 

•  Midipreps 

•  CD79A golden gate T4 ligation

•  Transformations
  •  CD79A golden gates 
  •  IgM RFP golden gate

•  3T3

•  6T6

•  6T12

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

^^^Pass: grapefruit

Non-Lab To Do's 

•  Cytometry time 
•  Order Cumate 
•  Chicken lysozyme, gp120
•  Antibodies for antigen binding detection

Waiting on primers to do list:

•  TRE:Syk-15-TEVp
•  CD79B-mRFP-G4 PCR

Higher Level To Do's:

•  Decide on other antigen-variable regions we want to test  
•  Is gantenerumab proprietary and can we actually publish with it?