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== CoRE MasterMix Preparation (total 20.168ul) == * Prepare the mastermix after the cells were ready to load # 2.33ul Phi29 10x buffer (from epicentre) # 8.45ul 200uM N6 primer # 1.688ul 25mM dNTP # 2.33ul 8% Tween20 # 2.11ul 1mM dUTP # 0.84ul Phi29 (1ug/ul) # 2.42ul water * Also prepare ALS and NS * ALS recipe: 69.5ul water + 10ul DTT(1M) + 8ul KOH(5M) + 12.5ul 8% Tween20 * NS recipe: 87.5ul NS stock + 12.5ul 8% Tween20 where NS stock: 4ml 1M HCl and 6ml 1M TrisHCl * Preparing "negative" control: added 0.35ul of water into 10ul CoRE mastermix in PCR tube. 15 hours at 30C. * Remaining mastermix volume was used to load the MDA device * All reagents were loaded onto the device as in the diagram below.
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