Monday's to Dos
- Midiprep
- Golden gates
- hEF1a:CD79B-pTet:RFP-Gal4VP16
- pENTR Syk-15-tTEV
- pENTR VRC01 light
- pENTR VRC01 heavy
- pENTR HyHEL-10 light
- pENTR HyHEL-10 heavy
- Pick Colonies for mini
- TRE:Syk-3-tTEV
- TRE:Syk-6-tTEV
- TRE:Syk-9-tTEV
- TRE:Syk-12-tTEV
- hEF1a:CD79A-9T15-G4
- Inoculate midis
- From failed midiprep
- Additional linkers
Tuesday's to Dos
- Transform golden gates
- hEF1a:CD79B-pTet:RFP-Gal4VP16
- pENTR Syk-15-tTEV
- pENTR VRC01 light
- pENTR VRC01 heavy
- pENTR HyHEL-10 light
- pENTR HyHEL-10 heavy
- Miniprep and digest
- TRE:Syk-3-tTEV
- TRE:Syk-6-tTEV
- TRE:Syk-9-tTEV
- TRE:Syk-12-tTEV
- hEF1a:CD79A-9T15-G4
- Midiprep and digest
- From yesterday's failed midipreps
- Additional linkers
- Pick colonies for failed linkers
- CD79A-3T15
- CD79A-12T9
- CD79A-15T9
- Send for sequencing
Wednesday's to Dos
- Redigest basically everything from yesterday
- Pick colonies
- TRE:Syk-15-tTEV
- hEF1a:CD79B-RFP-G4
- pENTR VRC01 heavy
- pENTR VRC01 light
- pENTR HyHEL-10 heavy
- pENTR HyHEL-10 light
- Figure out what to do about Syk linkers (possibly re-transform or re-plate?)
- Miniprep
- CD79A-3T15
- CD79A-12T9
- CD79A-15T9
- Inoculate midipreps if the sequencing comes back good
Sequencing account information:
^^^Email: bcrpeeps@mit.edu
^^^Pass: grapefruit