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===ALS and NS buffer preparation=== *Make fresh ALS buffer:(400mM KOH, 100mM DTT, 10mM EDTA) **Prepare 5M KOH: weigh one KOH pallet, add nuclease free H2O to make 5M KOH. Make fresh solution every week. **Mix 8 ul 5M KOH, 2 ul 0.5M EDTA, 10 ul 1M DTT, 80 ul nuclease free H2O **Mix 10ul ALS buffer with 10ul NS buffer, check pH with a pH paper, adjust ALS buffer with KOH or H2O if pH is not within 7-8 **Aliquot ALS buffer and NS buffer to two individual PCR tube, with 100 uL of each buffer. **UV treat NS and ALS buffer for 10 min.
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