Team:Goettingen/week16-3

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#3 Chemoreceptor Library - 16th Week

2^nd round: Mutagenesis PCR, 1000 µL

Experiment:
The PCR was set up in a volume of 1000 µL following the protocol from week 10.

V08_14 2^nd round: PCR purification

Experiment:
The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.

Observations & Results:
We now seemed to have found the mistake that caused problems with the loss of DNA material in the second mutagenesis round! Miscommunication led to using the wrong kit for purification steps. We decided to use the peqGOLD Cycle-pure Kit (PeqLab) instead of the subsequent ethanol precipitation as well, as this seemed to rise fewer problems.

V08_15 2^nd round: PCR purification

Experiment:
The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.

Observations & Results:
We now seemed to have found the mistake that caused problems with the loss of DNA material in the second mutagenesis round! Miscommunication led to using the wrong kit for purification steps. We decided to use the peqGOLD Cycle-pure Kit (PeqLab) instead of the subsequent ethanol precipitation as well, as this seemed to rise fewer problems.