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− | <img src="https://static.igem.org/mediawiki/2018/5/5b/T--munich--chi_inhibition_invitro.pdf " class="figure-img img-fluid rounded" alt=" | + | <img src="https://static.igem.org/mediawiki/2018/5/5b/T--munich--chi_inhibition_invitro.pdf " class="figure-img img-fluid rounded" alt="Chi invitro"> |
<figcaption class="figure-caption"></figcaption> | <figcaption class="figure-caption"></figcaption> | ||
</figure> | </figure> |
Revision as of 22:21, 17 October 2018
In Vitro inhibition of RecBCD with Chi6 Oligomers
2018/06/15Participants: | Dominic Schwarz |
Protocol: | Agarose gel, Restriction digest, Exonuclease V reaction | Notes: | Reaction was stoppedat at 10µl (50mM (2,82µl)) with EDTA (11mM), then heated on 70°C for 30 minutes. | Results: | Linear DNA was not degraded: we found out later that RecBCD was expired, but a positive control was missing. |
Redo: In Vitro inhibition of RecBCD with Chi6 Oligomers
2018/06/19Participants: | Enikö Baligács |
Protocol: | Agarose gel, Restriction digest, Exonuclease V reaction |
Notes: | The experiment was repeated by Enikö Baligács and a positive control was added. |
Results: | Linear DNA was not degraded: We found out that RecBCD was expired and not functional. |
Redo: In Vitro inhibition of RecBCD with Chi6 Oligomers
2018/07/02Participants: | Julia Mayer |
Protocol: | Agarose gel, Restriction digest, Exonuclease V reaction |
Notes: | No notes |
Results: | The Agarose gel was prepared wrong. Therefore, samples were lost and could not be evaluated. |
Linearizing pSB1C3_RFP
2018/08/01Participants: | Dominic Schwarz |
Protocol: | Restriction digest, PCR purification |
Notes: | EcoRI-HF |
Redo: In Vitro inhibition of RecBCD with Chi6 Oligomers. Digested backbone was stored.
2018/08/02Participants: | Enikö Baligács, Julia Mayer |
Protocol: | Agarose gel, Restriction digest, Exonuclease V reaction |
Notes: | The experiment was repeated with newly ordered recBCD |
Results: | Chi6 oligomers inhibit RecBCD but are degraded within 30 minutes. We went on to show these results in cell extract. |