In the first order derivative method, aliquots of MOX and CEF standard stock solutions were accurately transferred in to 10 mL volumetric flasks, separately and volumes were completed with water.
Separate quantities of the powder equivalent to 30 mg, 60 mg and 90 mg of PRG were transferred into a 100 mL volumetric flasks, dissolved in water, and sonicated for 5 min., the volume was then completed with water, shaken well for 5 min. and filtered into a dry flask.
A portion of the powder equivalent to 20.0 mg (method I), 40.0 mg (method II) and 30.0 mg (method III) of ALD was transferred into100-mL volumetric flasks, about 80 mL distilled water was added to each flask and they were sonicated for 15 min. The volume was completed with water, mixed and filtered.
A solution of F-FDG (70.6 and 70.5 MBq, as assessed by the dose calibrator) was introduced into a cylindrical phantom with an exactly known volume and completed with water, which resulted in a solution with an exactly known concentration.
The hybridization buffer contained 50%% of deionized formamide, 1.3× standard saline citrate (SSC; pH 5), 5 mM ethylenediaminetetraacetic acid (EDTA; pH 8.0; Sigma-Aldrich, Steinheim, Germany), 1 mg/ml of yeast tRNA (Sigma-Aldrich), 0.2 % Tween-20, 100 μg/ml of heparin (Sigma-Aldrich), completed with water (free of RNAase and DNAase; Sigma-Aldrich).
Apart from 200 U of M-MLV RT, the reaction mixture consisted of 250 pmol of random hexadeoxynucleotide primers (Roche Diagnostics GmbH, Penzberg, Germany), 4 μl of RT buffer (250 m M Tris-HCl pH 8.3, 375 m M KCl, 15 m M MgCl2, 50 m M DTT) and 4 μl of 1 m M dNTPs (Roche Diagnostics GmbH), completed with water to a final volume of 20 μl.
the solution completed with distilled water to the mark.
The homogenized matter is quantitatively transformed in a 250 mL measuring bottle and the volume is completed with distilled water.
The reaction mixture contained each primer at 0.5 μM, 200 μM deoxy nucleotide, Taq buffer, 2 U GoTaq Polymerase (Promega, Madison, WI, USA), 5 ng of DNA, completed with distilled water (Gibco) to 20 μL.
The rest of the volume was completed with distilled water.
Bags were weighed twice daily and the weight was completed with distilled water to maintain the humidity at 15%.
