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This mixture of solution was incubated at 37°C for 90 min and then the unconjugated free DNA was removed by the centrifugation.
Then, a mixture of solution of L-ascorbic acid (10%, 10 ml) and ginger extract (20 ml) was drop added to the silver nitrate solution under magnetic steering.
The following two solutions were prepared (concentrations given are those in the final CFPS mixture, which consisted of an equal mixture of Solution A and B): Solution A consisted of a small molecule substrate mixture [25] and 0.1% Tween 20 (Sigma Aldrich, St Louis, MO) to prevent bead aggregation.
The mobile phase was the mixture of solution A (water with 5 mM DBAA) and solution B (methanol with 5 mM DBAA) prepared at the gradient shown in Additional file 1: Table S3.
The solid support was transferred into a screw-capped Eppendorf tube and 1.5 mL of a 1 1 mixture of solution of methylamine in ethanol (33%%) ammonia and aqueous methylamine (40%%) was added and the reaction proceeded at room temperature for 4 to 6 h with occasional shaking.
This solution is a mixture of solution A: K4Fe CN 6.3H2O 5 m M, K3Fe CN 6 5 m M in wash buffer (MgCl2 2 m M, deoxycholate 0.01%, NP-40 0.02% in 0.1 M sodium phosphate buffer pH 7.8) and solution B: 5-bromo, 4-chloro, 3-indolyl β-d-galactopyranoside 50 mg ml−1 in dimethyl formamide) at ratio of 50 : 1.
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For the molecular size-selective detection of a protein formulation, a mixture of solutions of native SOD1 (100 μg/ml) and its aggregates (the volumetric ratio between them was 1 1, 1 5, 1 10, 1 20).
As surfactant and catalyst, 1 g polyvinyl pyrrolidone (PVP, average mol. wt. 100,000) and 0.5 ml HNO3 were added to the mixture of solutions containing Zn and Ta, respectively.
Once dried, the sections were rinsed twice for 2 min, and placed into a mixture of solutions D and E of the GolgiStain Kit for 10 min.
The mobile phase consisted of a variable mixture of solutions A (water-formic acid, 99 1 v/v) and B (acetonitrile formic acid, 99 1 v/v) at a flow rate of 0.3 ml min−1.
Fibrin glue was used according to the manual provided by the manufacturer in the form of a mixture of solutions A (fibrinogen and aprotinin) and B (thrombin and calcium chloride).
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