| Date | Summary |
|---|
| 06/01 | - h:Lyn-TEV Colonies Picked
- h:CDB-RFP-G4 transformation
- h:CDA-RFP-G4 Sequence Analysis
- Inocculate Midipreps
- h:Lyn, h:CDB-TCS-G4 cryostocks
|
| 06/02 | - Cloning Redesign
- Pick hEF1a:CD79B-pTet:RFP-Gal4VP16 Colonies
|
| 06/03 | - Miniprep hEF1a:CD79B-pTet:RFP-Gal4VP16
- Cryostock hEF1a:CD79A-pTet:RFP-Gal4VP16
|
| 06/04 | - Glycerol Stock hEF1a:CD79A-pTet:RFP-Gal4VP16
- Midiprep 4 Frozen pellets
- Digest & Gel hEF1a:CD79B-pTet:RFP-Gal4VP16
|
| 06/08 | - hEF1a:CD79B-pTet:RFP-Gal4VP16 Digest & gel
|
| 06/09 | |
| 06/11 | - Sequencing Analysis
- Glycerol Stock H:CDA-RFP-G4
|
| 06/16 | - designed SDM primers
- LR TRE:mKate, hEF1a:LacI, pLac:mKate
- experimental design + deadlines
|
| 06/17 | - annealed linker halves
- CD79A linker golden gates
|
| 06/18 | - transformed CD79A linker golden gates
- transformed LRs (TRE:mKate, pLac:mKate, hEF1a:LacI)
- LRed TREt:VP16Gal4
|
| 06/19 | - sdm primers are here
- killed LR TREt:VP16Gal4
- Transformed LR TREt:VP16Gal4
- Re-transformed CA-9T6-G4
- picked CD79A linker colonies
|
| 06/21 | - picked CA-9T6-G4 colonies
|
| 06/22 | - mini CD79A linkers
- mini TRE:mKate, pLac:mKate, hEF1a:LacI
- digest
- run CD79B SDM gel
|
| 06/23 | - digests and gels for TRE:mKate, pLac:mKate, hEF1a:LacI
- kill TRE:Gal4VP16 LR
- transform TRE:Gal4VP16 LR
- run SDM on gel -> gel extraction
- CD79B-RFP-G4 SDM product transformation
|
| 06/24 | - SDM CD79B-RFP-Gal4
- SDM DpnI digest
- transform SDM product
- transform TRE:Gal4VP16
|
| 06/25 | - redigest TRE:mKate, pLac:mKate, hEF1a:LacI
- PCR syk, tevp, ttevp
- gel verify PCRs and digested LRs
- TRE:gal4VP16 pick colonies and minipreps
- PCR purification of syk, tevp, ttevp
- reLR of TRE:mKate, pLac:mKate, hEF1a:LacI
- retransform SDM digests
|
| 06/26 | - TRE:gal4vp16 gel
- TRE:syk-mrfp-tevp golden gates
|
| 06/28 | |
| 06/29 | - cultured and miniprepped LR2 TRE:mKate, pLac:mKate, hEF1a:LacI
- miniprepped LR1 TRE:mKate, pLac:mKate, hEF1a:LacI
- digest and gel LR1 TRE:mKate, pLac:mKate, hEF1a:LacI
- picked colonies of new TRE:gal4vp16 transformation
|
| BCR 2.0 Cloning Progress 06/30 | - TRE:gal4vp16 miniprep, digest, gel
- hef1a:laci, tre:mkate digest, gel (new colonies with old enzymes)
- LR hef1a:cymR and cmv5CuO:mkate
- picked colonies for TRE:syk-mrfp-(t)tevp and pLac:mKate
|
| 07/01 | |