Cloning Strategy

CD79A Linker Optimization

Antibody Choices 

 Monday's to do's

•  

  Digest tTEVp and TEVp midis just to be sure (since there's an internal cut site)

•  Retransform
  •  B9T15 (from reaction)
•  Send for sequencing
  •  A3T15 #2 (8/15)

Sunday's to do's

•  Make amp plates
•  Replate
  •  B15T9
•  Inoculate miniprep cultures
  •  A15T9
  •  T:S-6-T
  •  B9T9
  •  B12T9
•  B15T9 Inoculate midiprep culture
  •  A9T15
  •  B6T9 probably B6T12
  •  B6T12 probably B6T9
•  Miniprep and sequence
  •  T:S-3-T
  •  B15T15
•  Spin down and store midiprep cultures
  •  T:S-9-tT
  •  T:S-12-tT
•  Analyze sequencing data
  •  B6T12
  •  B6T9
  •  A3T15

Saturday's to do's

•  Miniprep/sequence
  •  B15T15 - one red, one with no growth :/
  •  B6T12
  •  B6T9
  •  A3T15
•  Inoculate minis
  •  B9T9
  •  A15T9
  •  T:S-3-T
  •  T:S-6-T
  •  B12T9
  •  B15T15
•  Analyze sequencing and go from there

Friday's to do's

•  Midis
  •  B12T12-2
•  Minis, sequencing
  •  T:S-9-tT
  •  T:S-12-tT
•  Order antibodies/antigens
•  Finish Gaussian Distribution gating code
•  Split Cells
•  Inoculate mini
  •  B15T15
  •  B6T12
  •  B6T9
  •  A3T15
•  Hef1a:GAL4? 
•  Retransform 
  •  B9T9
  •  A15T9
  •  T:S-6-T
  •  B15T9
  •  B12T9
•  Sequencing, take 1
  •  B9T15-2
  •  B3T15-1
  •  T:S9tT 1
  •  T:S12tT 1
•  Sequencing, take 2
  •  T:S3T #2
  •  A9T15 #1 (8/12)

Thursday's to do's 

•  Midis:
  •  B9T12
  •  B15T3
  •  S9TEV
  •  S12TEV
•  Minis, sequencing 
  •  B6T9
  •  A9T15
•  Inoculate mini
  •  B15T9
  •  B15T15
  •  B12T9
  •  T:S-9-tT
  •  T:S-12-tT
•  Inoculate midi:
  •  B12T12-2
•  Sequencing:
  •  B6T9-1
  •  A9T15-2
  •  A3T15-2
  •  T:S-3-T 1
  •  T:S-6-T-1
•  Restreak:
  •  B15T9
  •  B12T9

Wednesday's to do's 

•  Midis:
  •  B9T12
  •  B15T3
  •  S9TEV
  •  S12TEV
  •  B12T12-2
    
•  Minis, sequencing:
  •  B15T9
  •  B15T15
  •  B12T9
  •  T:S-9-tT
  •  T:S-12-tT
•  Split cells 

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

^^^Pass: grapefruit

Check on status:

• B9T12 - send for seq
• B6T9 - send for seq
• B12T9 - repeat seq
• B12T12 - repeat seq

Midi's currently frozen in -80:

• B9T12
• B12T12
• B15T3
• S9TEV
• S12TEV
• S9tT
• S12tT

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