The following buffer solutions were tested: 2X SSC (300 mM NaCl, 30 mM sodium citrate), 2X SSPE (300 mM NaCl, 20 mM NaH2PO4, 2 mM EDTA), and, 2X STE (300 mM NaCl, 50 mM Tris-HCl, 2 mM EDTA).
For skin donors, skin samples, frozen in an 85% glycerol buffer, and skin storage solution were tested.
Local lesions typical of PMMoV infection were excised and crushed into 0.6 ml of inoculation buffer, and 350 µl of this solution were tested for the presence of PMMoV by RT-PCR using primers published by Hamada et al. [10], by electron microscopy, and by inoculation to one plant of Capsicum annuum var Yolo Wonder or Nicotiana benthamiana, two susceptible hosts for PMMoV.
To compensate for the removed 1.5 mL solution, 1.5 mL of fresh TRIS buffer solution was added to the original test solution.
At predetermined time intervals, 5 mL aliquots of the release medium were withdrawn from the release medium, and the same volume of fresh buffer solution was added to continue the release test.
At predetermined time intervals, 5 mL of the buffer solution was taken out for UV-VIS spectroscopy test and the concentration of the released drug was determined by recording the absorption spectra at 300 nm.
In a typical test, 0.5 mL of the drug containing buffer solution was removed and the vial was replenished with 0.5 mL of fresh buffer solution to maintain a constant volume of the released medium.
The buffer solution was phosphate buffer (6 mM, pH 7.4).
For this reason, 0.01 M of CuCl2 solution, 7.5 mM of ethanol neocuproine solution, and 1.0 M of CH3COONH4 buffer solution were added to each test tube, respectively.
For the test, 5 μl of serum or undiluted whole blood and 130 μl of buffer solution are added.
