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===Buffer Prep=== #1X Wash Buffer ##Warm 50X wash buffer to 40C for 10-20 minutes before mixing ##Add 2.94L DDI water and 60 mL 50X wash buffer to make 3 L 1X wash buffer ##1X wash buffer may be stored at RT for up to 1 month #50% Hematoxylin I ##Work in the fume hood ##Add 50% Hematoxylin by mixing hematoxylin I and DDI water ##Can be reused up to 1 week #0.02% Ammonia #Add 1.43 mL of 1N ammonium hydroxide to 250 mL DI water; mix well 3-5 times '''Do not reuse following reagents''' #XYLENE #70% and 100% EtOH used in Dehydration during counterstain
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