For extract without chlorophyll removal, methanolic extract of T. ornata and ethanolic extract of S. polycystum exhibited the highest DPPH radical scavenging activity.
For extract preparation, 1.5 kg of dried sample was extracted twice with 5 litre of 95% methanol at 25°C for 48 h.
For extract preparation, about 100 mg of powdered samples were added to 1.5 ml water, mixed in a plastic tube, incubated at 75°C for 20 min, and then centrifuged at 15,000 × g for 10 min.
For extract preparation we chose to use a strain of T. b. brucei that grows to relatively high densities in culture, even as bloodstream forms, since this is a prerequisite for large-scale cell fractionation and proteomic analysis.
For extract serial dilutions, plotting dilution factors versus relative response followed an inverse sigmoidal function, in which the apparent EC50 correspond to the dilution (actual or theoretical) giving 50% the response for 10 nM estradiol.
For extract preparation, tissue was ground to a white homogeneous powder in liquid nitrogen with 1 ml polysome buffer (10 mM Tris pH8, 150 mM NaCl, 5 mM MgCl2, 1% NP-40, 1% DOC, 10 mM DTT, 50 μg/ml cycloheximide, 0.4 U/μl RNAsin, 1 mM PMSF, 20 μg/ml aprotinin and leupeptin, supplemented with complete protease inhibitor (Roche Diagnostics, Indianapolis, IN, USA)).
For extract analysis, 6 spots of OT59 (1 μL/spot), 6 spots of P. citrinum (1 μL/spot), and 6 interactions of P. citrinum at a 5 mm distance from OT59 were inoculated on separate M1 agar plates (10 mL) and incubated at 30 °C for 48 h.
If nothing else, the book [see right for extract] means I finally have them all in one place".
Several protocols for extract purification were also studied.
Fungus Trichoderma virens ZY-01 (China patent ZL. 201210295819.6) was used for extract the total mRNA.
Top flowering parts were used for extract preparation (80% flowers and 20% rest of the plant).
