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Revision as of 23:58, 17 October 2018
Transforming E. Coli DH5a to amplify pKD3 for pRED/ET engineering
2018/05/28| Participants: | Dominic Schwarz |
| Protocol: | Electro-transformation |
| Notes: | No notes. |
| Results: | No colonies, no growth; We decided to use a resistance cassette from pSB1C3 without FRT sites. |
Amplifying a selection cassette from pSB1C3
2018/05/29| Participants: | Enikö Baligács |
| Protocol: | PCR, Agarose gel, Gel extraction |
| Notes: | PCR with two primer pairs were tested to amplify a chloramphenicol resistance cassette for the knockout. Both pairs have a 50 bp overhang homology region to the knockout target in E.Coli genomes. One pair has longer annealing sequence (CAT_RecBCD-KO_5HA_long_rv and CAT_RecBCD-KO_5HA_long_fw) and the other pair shorter. (CAT_RecBCD-KO_5HA_short_rv and CAT_RecBCD-KO_5HA_short_fw). |
| Results: | We thought the band might be what we wanted but because of how few DNA the extraction yielded, these samples were not used in further experiments.
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