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Extraction steps were undertaken following the protocols of the manufacturer, apart from the DNA elution step, where all samples were eluted with 50 μL elution solution.
However, a lower second elution step could also result in co-eluting further contaminants.
P34 could be eluted from the column during the second elution step (0.4 M (NH4 2SO4).
GST-GraS and GraX co-eluted in the first fraction of the elution step.
This elution step was repeated 5 times and the elutant and beads were stored at −20°C.
For the elution step, use sterilized MQ water (50 μl) to elute the template plasmid.
Since this elution step specifically disrupts the Gαi1-Gβγ interaction, the eluted fraction is fairly pure.
The elution step was repeated a second time and the volume of the eluted peptides were brought to 500 μL by SpeedVac concentration.
The elution step is carried out with 1.0 mol L−1 HNO3.
Binding buffer was added 13x to the concentrated supernatant and DNA was purified with MinElute columns (Qiagen), following the manufacturer's instructions with the exception of a 15-minute incubation at 37 °C during the elution step.
The N2 gas is used to removal solvents in elution step.
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