Team:Munich/chiinhibition1.html

In Vitro Inhibition of RecBCD with Chi6 Oligomers

2018/06/15
Participants: 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

2018/06/19
Participants: 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

2018/07/02
Participants: 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

2018/08/01
Participants: Dominic Schwarz
Protocol: Restriction digest, PCR purification
Notes: EcoRI-HF

Redo: In Vitro Inhibition of RecBCD with Chi6 Oligomers

2018/08/02
Participants: Enikö Baligács, Julia Mayer
Protocol: Agarose gel, Restriction digest, Exonuclease V reaction
Notes: The experiment was repeated with newly ordered recBCD.
Digested Backbone was Stored.
Results: Chi6 oligomers inhibit RecBCD but are degraded within 30 minutes. We went on to show these results in cell extract.
Chi invitro