BCR 2.0

Cloning Strategy

CD79A Linker Optimization

Antibody Choices 

Wednesday's To Do's

• Run gel pENTR HyHEL10 H
• Transform 
  • B15T15
• Mini, sequence
  • A9T15 
  • B9T9 
• Midi:
  • TRE:Syk-15-tTp
• inoculate midi:
  • B15T3
  • T:S-9-TEVp
  • T:S-12-TEVp
  • B9T12
• inoculate mini
  • A9T15
  • B15T9
  • B6T9
• seq.
  • B6T12 2
  • pENTR HyHEL10 H 1
  • A15T9 2
  • B9T9 (8/12)
• repeat seq.
  • T:S-3-T 1
  • T:S-6-T 1
  • T:S-9-tT 2
  • T:S-12-tT 2
  • B3T15 1
  • B9T15 1
  • B12T12 2

Tuesday's To Do's 

• Gel TRE:Syk-15-tTEVP -> send for sequencing 
• digest, gel, pENTR HyHEL10 H -> send for sequencing 
• sequencing:
  • B6T9-1
  • B9T12-1
• innoculate midi:
  • B15T3
  • T:S-15-tT

• inoculate mini:
  • B9T9 - from plate
  • A3T15 - from plate 
• restreak/replate
  • B15T9
  • A9T15 - no white
  • B15T15 
  • B9T9 - only one white

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

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