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Monday's to Dos

  • Midiprep
    • something
  • Golden gates
    • hEF1a:CD79B-pTet:RFP-Gal4VP16
    • pENTR Syk-15-tTEV
    • pENTR VRC01 light
    • pENTR VRC01 heavy
    • pENTR HyHEL-10 light
    • pENTR HyHEL-10 heavy
  • Pick Colonies for mini
    • TRE:Syk-3-tTEV
    • TRE:Syk-6-tTEV
    • TRE:Syk-9-tTEV
    • TRE:Syk-12-tTEV
    • hEF1a:CD79A-9T15-G4
  • Inoculate midis
    • From failed midiprep
      • hEF1a:Gal4
      • GGDonr
      • Linkers
    • Additional linkers
      • CD79A-6T9
      • CD79A-9T12

Tuesday's to Dos

  • Transform golden gates
    • hEF1a:CD79B-pTet:RFP-Gal4VP16
    • pENTR Syk-15-tTEV
    • pENTR VRC01 light
    • pENTR VRC01 heavy
    • pENTR HyHEL-10 light
    • pENTR HyHEL-10 heavy
  • Miniprep and digest
    • TRE:Syk-3-tTEV
    • TRE:Syk-6-tTEV
    • TRE:Syk-9-tTEV
    • TRE:Syk-12-tTEV
    • hEF1a:CD79A-9T15-G4
  • Midiprep and digest
    • From yesterday's failed midipreps
      • hEF1a:Gal4
      • GGDonr
      • Linkers
    • Additional linkers
      • CD79A-6T9
      • CD79A-9T12
  • Pick colonies for failed linkers
    • CD79A-3T15
    • CD79A-12T9
    • CD79A-15T9
  • Send for sequencing
    • Resend 12T12 (1), also send 12T12 (2)
    • 9T15 (1) and (2)

Tuesday's to Dos

  • Transform golden gates
    • hEF1a:CD79B-pTet:RFP-Gal4VP16
    • pENTR Syk-15-tTEV
    • pENTR VRC01 light
    • pENTR VRC01 heavy
    • pENTR HyHEL-10 light
    • pENTR HyHEL-10 heavy
  • Miniprep and digest
    • TRE:Syk-3-tTEV
    • TRE:Syk-6-tTEV
    • TRE:Syk-9-tTEV
    • TRE:Syk-12-tTEV
    • hEF1a:CD79A-9T15-G4
  • Midiprep and digest
    • From yesterday's failed midipreps
      • hEF1a:Gal4
      • GGDonr
      • Linkers
    • Additional linkers
      • CD79A-6T9
      • CD79A-9T12
  • Inoculate midiprep cultures for successful CD79A linkers (based on sequencing data that we should get back today) and pick new colonies for not successful ones



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:
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?

THINGS TO NOT FORGET TO DO:

  • SYK-15tTEVP golden gate when we figure out what's wrong with the AarI
  • 3T3 MIDI

 

 

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