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==Sample & Material== *Primer:L4N6 primer *Cell: GM18506 cell line, harvested, washed by UV-treated PBS and coundted to 200,000 cell/mL (200 cell/uL) **Dilute to 10 cell/uL, 5 cell/uL, 0.5 cell/uL using UV-treated PBS Dilute factor original x 1/2 x 1/10 x 1/2 x 1/10 conc.(cell/uL) 200/uL 100/uL 10/uL 5/uL 0.5/uL Transfering(uL) 50 10 50 10 UVed-PBS(uL) 50 90 50 90 *Pos. control: human gDNA diluted to 3 ng/uL **Measure the original (commercial human gDNA) using Nanodrop -> 309ng/uL Dilute factor original x 1/100 x 1/10 x 1/10 x 1/10 conc. 309ng/uL 3ng/uL 300pg/uL 30pg/uL 3pg/uL Transfering(uL) 1 10 10 10 UVed-PBS(uL) 99 90 90 90 *Make fresh ALS buffer:(400mM KOH, 100mM DTT, 10mM EDTA) **Prepare 5M KOH: weigh one KOH pallet in a 1.5mL-tube. Add nuclease-free H2O to make 5M KOH. **Mix 20 ul 5M KOH, 5 ul 0.5M EDTA, 25 ul 1M DTT(Qiagen), 200 ul nuclease free H2O '''(Note: this is new formula)''' **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 0.093 g (KOH=56.11) = 0.001657 mole; 0.001657/5 x 10^6 =331.49 uL --- H2O needed
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