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Firstly, each sample was soaked by aether for 10 min to remove proteins and fattiness on the samples' surface.
Subsequent treatment with RNase (1 mg/ml) and phenol chloroform:isoamylalcohol (25:24:1) was undertaken in order to remove proteins.
Regarding studies with sole surfactant treatment, Kim et al. (2003) applied SDS to Ralstonia eutropha cells, but additional heating at 121°C and washing steps were required to remove proteins and achieve a final purity of 97%.
When researchers do this, they also remove proteins that are essential to help cells divide, Lanza said.
Urine was lyophilized before MRS, while serum was analyzed following ultrafiltration to remove proteins.
Alternatively, a filtration step to physically remove residual dye would also work, but it might also unspecifically remove proteins.
Two extractions with phenol-chloroform-isoamyl alcohol (25∶24∶1) and one with chloroform were used to remove proteins.
The initial set of proteins was limited to those that could be confidently identified, and was further screened to remove proteins with few non-zero peptide hits.
Before usage in this study, DNA aliquots were re-precipitated and treated with RNAse to remove proteins and RNA remnants (Fig. S1).
The methodology we employed to remove proteins from the surface of USA300 was not specific to conventionally defined surface proteins (i.e., cell wall-associated and membrane proteins).
S. solfataricus cell lysate was incubated with the immobilized DPSL followed by several steps of washing to remove proteins that bound nonspecifically.
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