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EricChu:RepliPHI MDA protocol (20ul reaction)
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==RephiPHI MDA Protocol (20ul reaction)== #Start with 1.0ul Ξ» DNA (2.5ng/ul) in PCR tube #Add 1.5ul ALS, incubate at room temp 3mins #Add 1.5ul NS, put on ice immediately #Create master mix (w/ 1% Tween20) ##2ul 10x buffer ##5ul 200uM N6 primers ##0.8ul 25mM dNTPs ##2ul Phi29 polymerase ##2ul 8% Tween20 ##4.2ul H20 #Add master mix to PCR tube #Add to thermocycler for 10 hours at 30β°C and keep at 4 β°C #Use PCR purification kit to collect DNA #Measure DNA concentration with NanoDrop ALS: 69.5ul H20 + 10ul DTT(1M) + 8ul KOH (5M) + 12.5ul 8% Tween20 NS: 87.5ul NS stock + 12.5ul 8% Tween20 where NS stock: 4ml 1MHCl and 6ml 1MTrisHCl
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