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===Isolate RNA from BA8 tissue section=== *Using ZR RNA MicroPrep kit from Zymo #Wipe all surfaces, forceps, etc with EtOH and RNase Zap #Scrape tissue off glass with a scalpel and put in 1.5ml tube #Add 400ul RNA Lysis Buffer #*Vortex 10sec and pipette with 1000p ~10x #Centrifuge at 18,000rcf for 1min #Transfer the 400ul to IIIC column and centrifuge at 8,000rcf for 30sec #Add 320ul 100% EtOH (UV'd) to flow-trhough and mix #Transfer 720ul to IC column and centrifuge at 18,000rcf for 1min #Add 400ul RNA Wash Buffer and centrifuge at 18,000rcf for 30sec #Add 30ul DNase I cocktail and incubate in 37C incubator for 15min #*1.5ul DNase I (2U/ul) + 3ul 10X Reaction Buffer + 25.5ul RNA Wash Buffer #*Centrifuge at 18,000rcf for 30sec #Add 400ul RNA Prep Buffer and centrifuge at 18,000rcf for 1min #Add 800ul RNA Wash Buffer and centrifuge at 18,000rcf for 30sec and repeat with 400ul Wash Buffer #Spin in emptied collection tube at 18,000rcf for 2min #Add 16ul H2O, let stand for 1min, and then centrifuge at 10,000rcf for 30sec *Nanodrop Measurement: 6.9ng/ul **A260/280 = 1.77
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