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 minutes 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 minutes 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 minutes 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 minutes 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 minutes 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 minutes 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: VF2 & VR. |
| Results: | The Agarose gel after PCR showed no correct bands, the template seems to be incorrect. |