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
  • B12T9
  • B15T9
• Inoculate miniprep cultures
  • A15T9
  • T:S-6-T
  • B9T9
• 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