Today's To Do's:
- pour kan x-gal plates
- restreak TRE:Syk-mRFP-(t)TEVp
- pick TRE:Syk-mRFP-(t)TEVp
- miniprep digest verify TRE:Syk-mRFP-(t)TEVp
- transform pENTR LacI and pENTR pLac
- pick hEF1a:LacI, TRE:mKate, pLac:mKate
- digest, gel hEF1a:LacI, TRE:mKate, pLac:mKate LR2
- miniprep TRE:gal4 transformation 2
- digest, gel verify TRE gal4 transformation 2
- split cells
- pLac:mKate pick colonies
- ScaI to lab stock
Geneious To Do List:
SDM primers: CD79B, Gal4VP16, TEVp
- TRE:gal4vp16 sequencing primers
- TRE:Syk-mRFP-(t)TEVp sequening primers
- 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