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===PCR with Amp6.3NH2 primer=== I did the PCR reamplification with Amp6.3NH2 primers. 400ul for each sample. reaction system x4 x2 H2O 45ul 180ul 360 2x iProof Mastermix 50ul 200ul 400 AmpF6.3NH2(10uM) 2ul 8ul 16 AmpR6.3NH2(10uM) 2ul 8ul 16 50x SYBG I 0.4ul 1.6ul 3.2 template 1ul/each for purified amplicons Total 100ul 800ul 98C 30S -> (98C 10S -> 58C 20S -> 72C 20S) x 8 ->72C 3min. Purify with Qiaquick column. Elute in 100ul EB.
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