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Daniel:Notebook/RNAFACS/2015-11-20
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=PAGE Gel Creation= [[Daniel:Notebook/RNAFACS|Back to Calendar]] The reagents finally came in for making gels on slides. Today I'll start with protocol #1. ==Day 1-Silane Binding== #Load slides (facing the same direction) into metal slide racks #Sterilize the following items (15 minutes under UV light in PCR hood): ##two large glass Coplin buckets & lids ##metal slide racks with slides loaded ##1 L dH20 in a plastic container ##stir bar #Add 220 uL Acetic Acid to the 1 L dH20 (pH -> 3.5; no need to measure) #Add 4 mL Bind-Silane #Mix for 15 minutes using stir-bar. #Place loaded slide racks in glass Coplin buckets #Add stirred mixture to Coplin buckets (500 mL per bucket; enough to cover the slides) #Cover buckets and incubate with slow shaking for 1 hour. #Leaving racks in buckets, rinse 3 times with dH20 #Rinse once with 100% ethanol. #Remove slide-racks from buckets and allow slides to dry (in racks) under AirClean hood. #Transfer slides to plastic storage boxes (keep the Teflon coated sides of the slides facing in the same direction) #Label boxes with initals and date of treatment. #Place slide boxes in dessicator for long-term storage.
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