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In the simulation, we tested the unstable accuracy of the analytical estimation, the consistent accuracy of the maximum sample size determined by the search algorithm and the advantages of the proposed design on saving sample size. Cancer stem cells (CSCs) are considered the main cause of cancer recurrence and metastasis, and DNA methylation is involved in the maintenance of CSCs. However, the methylation profile of esophageal CSCs remains unknown. Side population (SP) cells were isolated from esophageal squamous cell carcinoma (ESCC)
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