Team:Munich/recbcdtev.html
PCR to generate RecBCD with engineered TEV sites
2018/07/27| Participants: | Enikö Baligács |
| Protocol: | PCR, Agarose gel |
| Notes: | RecBD:RecB-TEV-fw & BBs-PstIrv; Template: Mini pSB1C3_lacO_RecBCD-His(1) (unconfirmed);
AT: 57° ET: 2 min 30 s (2,7 kb) RecCB: RecB-TEV-rw & RecC-TEV-fw; Template: Mini pSB1C3_lacO_RecBCD-His (1) (unconfirmed); AT: 61,5 ° ET: 3 min 30 s (3,7 kb) TEV-RecC-lacO-BBP: RecC-TEV-rw & BBP_EcoRI_fw; Template: Mini pSB1C3_lacO_RecBCD-His (1) (unconfirmed); AT: 61,5 ° ET: 3 min 30 s (2,9 kb) |
| Results: | no bands. We decided to redo the PCR reaction. |
Redo: PCR to generate RecBCD with engineered TEV sites
2018/07/29| Participants: | Enikö Baligács |
| Protocol: | PCR, Agarose gel |
| Notes: | RecBD:RecB-TEV-fw & BBs-PstIrv; Template: Mini pSB1C3_lacO_RecBCD-His(1) (unconfirmed);
AT: 57° ET: 2 min 30 s (2,7 kb) RecCB: RecB-TEV-rw & RecC-TEV-fw; Template: Mini pSB1C3_lacO_RecBCD-His (1) (unconfirmed); AT: 61,5 ° ET: 3 min 30 s (3,7 kb) TEV-RecC-lacO-BBP: RecC-TEV-rw & BBP_EcoRI_fw; Template: Mini pSB1C3_lacO_RecBCD-His (1) (unconfirmed); AT: 61,5 ° ET: 3 min 30 s (2,9 kb) |
| Results: | Did not work again. We suspected that the template is incorrect. |
test digest of pSB1C3_recBCD(WT)
2018/07/30| Participants: | Dominic Schwarz |
| Protocol: | Restriction digest, Agarose gel |
| Notes: | PstI & EcoRI |
| Results: | The agarose gel showed no correct bands, the template is incorrect. |
Test PCR of recBCD WT
2018/07/27| Participants: | EniKÖ Baligács |
| Protocol: | PCR, Agarose gel |
| Notes: | Primers: |
| Results: | The agarose gel after PCR showed no correct bands, the template seems to be incorrect. |