Kun:LabNotes/ExonomeSeq/2007-12-17
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Exp.1 Start another round of E55kv1 probe production
PCR (with NEB Taq master mix)
x1 x96 E55k1(20nM) 0.1ul 9.6ul 2x Taq master mix 50ul 4800ul 100uM pAP1V6U 0.3ul 28.8ul 100uM phosAP2V6 0.3ul 28.8ul H2O 49.3ul 4800ul
94C 3min -> 18 cycles of (94C 45sec -> 58C 2min -> 72C 1min) -> 72C 3min -> 15C hold
Ethanol precipitation. I put everything in a 50-ml tube for centrifugation. The tube broke during spinning. And I lost ~30% of the sample. The remaining sample contains some debris. After resuspending with 200ul ddH2O, I filtered the solution with NanoSep column.
The Nanodrop read is 4.3ug/ul, but it might not be accurate. I did lambda exo digestion on half of the DNA: 100ul DNA + 60ul H2O + 20ul 10X buffer + 20ul lambda exo -> 37C 8h -> 85C 20min -> 4C hold.