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==2nd PCR amplification with AmpF6.3/AmpR6.3 and dUTP:dNTP 1:40 == reaction system x4 x3 H2O 43.2ul 172.8ul 518.4 2x Master mix 50ul 200ul 600 dUTP(1mM) 2ul 8ul 24 AmpF6.3(10uM) 2ul 8ul 24 AmpR6.3(10uM) 2ul 8ul 24 50x SYBG I 0.4ul 1.6ul 4.8 template(18.4ng/ul) 0.1ul 0.4ul 1.2 Total 100ul 400ul 2400 94C 3min -> 13cycles of (94C 45sec -> 55C 45sec -> 72C 45sec) -> 72C 3min -> 4C Bioneer column purification. Yield:167ng/ul x60ul
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