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===PCR=== PCR x 2 Template: 10ul 2X iProof MasterMix 50ul 100uM AmpF6.3rSol 0.4ul 100uM AmpR6.3rSol 0.4ul 50X SYBG I: 0.4ul H2O 38ul 98C 30sec -> (98C 10sec -> 60C 20sec -> 72C 15sec) x 8 -> (98C 10sec -> 72C 20sec) x 12 -> 72C 3min -> 4C hold. *The amplification curves came close to plateau at 12th cycles. The capturing seems to worked for both reactions. So I repeat the amplification on the remaining 10ul capturing reactions. *I got 200ul amplicons for each sample. I did E-gel SizeSelection on 120ul amplicons. It didn't go very well because of (i) high salt concentration in the PCR buffer; (ii) too much DNA. Nonetheless, the size-selected DNA were purified with Qiaquick columns together with the remaining 80ul amplicons. *I did a quantitative PAGE gel on all the four DNA samples. The bands are fuzzy due to the presence of ethanol in the eluted DNA. *Based on the PAGE gel, the estimated concentrations are: CES8k18bp-Hues6-DSCp: 4.5ng/ul = 38nM CES8k18bp-Hues6-tRp: 3.3ng/ul = 28nM
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