Today's To Do's:
- digest TRE:Gal4VP16
- pick hEF1a:LacI, TRE:mKate, pLac:mKate colonies
- kill LRs
- syk-rfp-tvep/ttvep linker golden gates (cidar and overnight)
- gel verify TRE:Gal4VP16 digest
- miniprep hEF1a:LacI, TRE:mKate, pLac:mKate colonies
- transform LRs
- transform golden gates
- transform pENTR LacI and pENTR pLac
- order new SDM primers
- send TRE:Gal4VP16 for sequencing
Other Immediate To Do's:
- Innoculate TRE:mKate midi culture (culture number 2)
- transform golden gates
- get cells from Jeremy
^^^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