After this purification phase, blood is reconstituted and dialyzed with standard techniques.
The purification of the lysate was performed with a slight modification from the protocol in which magnetic particles (MagAttract® Suspension G, QIAGEN) were used to shift DNA through purification phases using a Kingfisher (ThermoScientific) automated purification system.
Total RNA was extracted from nasal mucosa samples using RNeasy Mini Kit (Qiagen, Valencia, CA, USA) with the addition of a DNAse treatment (Promega, Madison, WI, USA) and re-purification phase.
The purification procedures involving solid phase extraction (SPE) based methods with both commercial Isolute-Florisil cartridges, and purpose designed molecularly imprinted polymeric materials (MIPs), resulted in cleaner chromatograms, while the mycocerosates are still present.
Both are compared against the unmutated sequence, designated Z4, which is currently utilized in a commercially available Protein A stationary phase for the purification of molecules containing Fc domains.
The positive effect of variable B and the negative effect of A and of the interaction between A and B on the purification factor in the top phase (PFT) indicated that a simultaneous increase in PEG concentration with a decrease in molar mass of PEG leads to an increase in that response.
The purification step with the solid-phase anion-exchange column presumably renders the assay free of interferences.
Finally, our biochemical approach based on the purification of PM by two-phase PEG/dextran partitioning clearly demonstrated the presence of HA-CaNDR1a proteins in tobacco PM fractions.
Response surface methodology was used to determine an optimized aqueous two-phase system for the purification of rGUS from transgenic tobacco.
The PEG3350, citrate and sodium chloride concentrations were selected as variables to evaluate the purification impact factors in aqueous two-phase system, including partition coefficients of α-amylase, total protein, purification factor and α-amylase yield.
