Wednesday's To Do's
- Flow cytometry with Brian
- Midiprep (and verify?)
- pEXPR TRE:Syk
- hEF1a:CymR
- hEF1a-CuO:mKate
- CMV5-CuO:mKate
- PCRs of mRFP and mIgM constant region for hEF1a:mRFP-CH (constant heavy chain)
- Order chicken lysozyme and gp120?
- LR - probably can't do today because no LR clonase
- EGSH:mKate
- hEf1a:VgEcR-2A-RXR
- hEF1a:mKate
- hEF1a:Gal4VP16 (remember which pDEST)
- hEF1a:CD79B-TCS-Gal4VP16
Non-lab to do's
- Design (or find?) sequencing primers for mIgM CH (constant region)
- Plan experiment to test gantenerumab binding to beta-amyloid
- Leader sequences for ordering gBlocks of variable regions?
- Review sequencing
- TRE:Syk-pTet:mRFP-TEV - priming issue
- TRE:Syk-pTet:mRFP-tTEV - priming issue
- pENTR CD79B-TCS-G4 - correct
CMV5-CuO 715:
hEF1a-CuO 715:09
CymR-CuO 710:09
Geneious To Do List:
- Antibody cloning for chicken lysozyme - Kathryn
hef1a sequencing primers
^^^Email: bcrpeeps@mit.edu
^^^Pass: grapefruit
Higher Level To Do's:
- Decide on other antigen-variable regions we want to test
- Other experimental planning (on experimental page)
- Parts list (excel sheet detailing specs/locations of parts)
- How do we do site directed mutagenesis?
- Literature research:
- kozak sequences for heavy and light chains and CD79s
- truncated TEVp
- other antigens to detect using BCR (we were thinking of small molecule detection)
- inducible promoters (we were considering LacI - an inducible repressor - but we want to look into other options first)
- Cloning planning
- hEF1a:Syk-pTet:mRFP-TEVp
- variable region
- Is gantenerumab proprietary and can we actually publish with it?