Difference between revisions of "Team:Munich/deletioncasette.html"
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| − | <h4>Transforming E. Coli DH5a to | + | <h4>Transforming E. Coli DH5a to Amplify pKD3 for pRED/ET Engineering</h4> |
<em>2018/05/28</em> | <em>2018/05/28</em> | ||
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| − | <h4>Amplifying a | + | <h4>Amplifying a Selection Cassette from pSB1C3</h4> |
<em>2018/05/29</em> | <em>2018/05/29</em> | ||
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
Latest revision as of 00:13, 18 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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