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After the resulting solution was maintained at −78 °C for 30 min, a solution of 15 (2.00 mmol, 1.0 equiv) in THF (1 mL) was slowly added for 10 h.
After stirring the reaction mixture for 10 min, a solution of 1,2-di-toluenesulphonyloxyethane (1.85 g, 5 mmol) in 10 mL of THF was added, and the reaction mixture was refluxed overnight.
Definition 1: In the molecule emission control bargaining game ( W,U min), a solution (mathbf U^{ast }=left (U^{ast }_{1},U^{ast }_{2}right)) is said to be an NBS in W if the following axioms are satisfied.
After stirring the reaction mixture for 10 min, a solution of 1-fluoro-2-tosyloxyethane (1.09 g, 5 mmol) in 10 mL of tetrahydrofuran was added, and the reaction mixture was refluxed overnight.
After 30 min, a solution containing 4- 2-methoxyethoxy methoxy -3-methoxy 4- 2-methoxyethoxy methoxy -3-methoxy 4- 2-methoxyethoxy methoxy -3-methoxy 4- 2-methoxyethoxy methoxy -3-methoxy 4- 2-methoxyethoxy methoxy -3-methoxy
After 15 min, a solution of 3-azido-propionaldehyde 10 in THF was added dropwise at −78 °C.
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The slide was placed on dry-ice for 20 min, the cover slip removed and then washed twice in PBS for 10 min each and blocked for 30 min in a solution of PBS containing bovine serum albumin (BSA).
Cells were washed 3 times (5 mins).in PBS and incubated for 20-25 min in a solution containing 10% horse serum, 0.125% TritonX-100 in PBS.
Gels were sensitized for 45 min in 50 ppm sodium thiosulfate solution, soaked in 0.1% silver nitrate for 45 min, rinsed with ultrapure water, and developed for approximately 1 min in a solution of 2% sodium carbonate with 0.037% formaldehyde.
The reaction was kept at −78°C for 15 min, then warmed to room temperature and stirred for a further 30 min. A solution of Na2SO310%% in water (1.2 mL) was added to destroy the excess of peroxide.
For ACF and tumor measurements, colons were stained for 5 min in a solution of 1% methylene blue in PBS and destained for 5 min in PBS.
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