• Created by Unknown User (lylaatta@mit.edu), last modified by Unknown User (clrichar@mit.edu) on May 28, 2015 15:06

You are viewing an old version of this page. View the current version.

Compare with Current View Page History

« Previous Version 41 Next »

BCR 2.0 

Experimental Progress:

Cloning Progress
Parts Tree
Cloning Strategy
Experiments
Linker Optimization
Antibody Choices 

 

Useful Links: 

Protocols
Immediate To Do's:
TaskClaimed byDate CompletedOther Notes
CD79A/B, Gal4VP16, RFP PCR try3CRApr 10

 

Gel verification of try3 PCR productLAApr 10RFP and Gal4 worked, CD79s didn't - we probably need to redesign primers
Redesign primersLA + CRApr 11CD79A F and R and CD79B F primers were redesigned and ordered
AarI golden gate of CD79A constructLAApr 30 
Take out CD79A golden gate platesLAMay 2 

Make LB

CRMay 4 

CD79A golden gate liquid cultures

LAMay 4

Cultures in at 6pm - need to be taken out by 10/11 am May 5

in 215 shaking incubator

Take out CD79A golden gate liquid culturesLAMay 5In 4C

CD79A golden gate minipreps

LAMay 6In yellow rack in 4C - need to be nanodropped

CD79A golden gate restriction verification

CRMay 6Some promising constructs
Send promising CD79A golden gates for sequencing   
  • CD79A linker golden gates
   
Redesign CD79B reverse PCR primerCRMay 24 
TaskClaimed byDate CompletedOther Notes
Order primers for PCRing constant region  primers on geneious
    

Christian is Confused so he is using this to do list

Day

List

Monday

  • None

Tuesday

  • Design CD79B Sequencing Primers
  • Look into tTEV and IGD options
  • Apply for SynBerc and BDAC
  • Plan Lyn and Syk Experiment
  • Plan mAB cloning

Wednesday

  • Establish HEK293 Cell Line
  • Establish Cryostocks of Gmab H, Gmab L, Syk-TEV, Lyn-TEV, CD79A, CD79B, CD79A-G4, CD79B-G4, hEF1a: CD79A - pTet: RFP - Gal4VP16
  • Transfect Lyn/TEV experiment

Thursday

  • Sequence hEF1a: CD79A - pTet: RFP - Gal4VP16
  • PCR CD79B with third gen primers
  • Digest & Gel CD79B &hEF1a: CD79A - pTet: RFP - Gal4VP16
  • GG hEF1a:CD79B-pTet:RFP-Gal4VP16
  • GG H:CDA-RFP-G4 with linkers

Friday

  • 9:00 AM Cytometry
  • Transform GG hEF1a:CD79B-pTet:RFP-Gal4VP16
  • Transform H:CDA-RFP-G4 w/ linkers

Saturday

  • Pick Colonies hEF1a:CD79B-pTet:RFP-Gal4VP16
  • Pick colonies H:CDA-RFP-G4 w/ linkers
  • Split HEK293

Sunday

  • Miniprep hEF1a:CD79B-pTet:RFP-Gal4VP16
  • Miniprep H:CDA-RFP-G4 W/ Linkers
  • Digest & Gel hEF1a:CD79B-pTet:RFP-Gal4VP16
  • Digest & Gel H:CDA-RFP-G4 W/ Linkers

Day

List

Monday

 

Tuesday

 

Wednesday

 

Thursday

  • Transform For Cryostocks, Midipreps, Minipreps (Including Lyn)
  • Split HEK Cells
  • Sequence H:CDA-RFP-G4
  • PCR CD79B with 3rd Gen primers
  • Gel CD79B
  • GG H:CDB-RFP-G4

Friday

  • Transform H:CDB-RFP-G4
  • GG H:CDA-RFP-G4 With Linkers
  • Pick Colonies for Midipreps/Cryostocks

Saturday

  • Midiprep new DNA stocks
  • Create duplicate cryostocks
  • Pick colonies for H:CDB-RFP-G4
  • Transform GG H:CDA-RFP-G4 w/ linkers

Sunday

 
Higher Level To Do's:
  • Finish Discussing Figures for Experiment list
  • Decide on other antigen-variable regions we want to test 
  • Syk-Lyn experiment design 
  • Receptor dilution experiment design
  • Other experimental planning

 

  • No labels