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== Sample list in each step in brief == ==== Cell lysis ==== :- WT_1 to WT_16 (WT 1 nu, performed by Youjin). I re-run the number of samples to make it easy to handle and track. :For my part I will include all controls and also in the following steps :- T_1 to T_16 (TKO 1 nu, performed by Noi). :- T10, T100 and T0 (TKO 10nu, 100nu and 0nu, 1 each, performed by Noi) :- 10pg of CTT with unmeth lambda DNA (duplicates). CTT is gDNA from frozen colon tumor tissue purchased from Biochain. :- NTC with unmeth lambda DNA (duplicate), * All samples in this step will have 60fg of unmeth lambda DNA spike in. This allows for bisulfite conversion efficiency measurement. :- I did serial dilution of CTT gDNA from 2ng/ul to 100pg/ul and 5pg/ul ::- Mix 10ul of 2ng/ul CTT gDNA with 190ul H2O ::- Mix 10ul of 100pg/ul CTT gDNA with 190ul H2O ==== DNA fragmentation with MspI ==== :- 10pg of CTT '''without''' unmeth lambda DNA (duplicates). :- MspI NTC '''without''' unmeth lambda DNA (duplicates). ==== Methylated adaptor ligation ==== :- Ligation control (duplicates). * No NTC during Gap-filling/dA-tailing
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