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==Running Real-time MDA in PCR tubes to test different MDA conditions == * Use both PGP1 cells and genomic human DNA # 1ul 400mM ALS was added to 100 cells in 1ul PBS. Incubated for 1min in room temp. # 1ul 400mM NS was added. Incubated for 1min in room temp. # Added 1ul of (i) water (no protease) or (ii) 24mM protease and incubate at 40C or 50C for 15min # Added 1ul of 250mM KOH (final 50mM) or 100mM KOH (final 20mM) # Added 5ul of 50mM HCl or 20mM HCl (according to the previous step). Also added 5.5ul HCl (10% extra) and 6ul HCl (20% extra) to the test. # Took 1ul out of the 10ul from above (equivalent to 10 cells) and added to 40ul MDA mix with SYBR Green.
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