DNA was isolated using a Qiagen DNA isolation kit, then COBRA using the bisulfite conversion kit (Qiagen) was performed.
DNA was isolated from human NSCs, ESCs and iPS cells and treated with bisulfite using the EpiTect Bisulfite Kit (Qiagen) according to manufacturer's instructions.
Genomic DNA (1 µg, or 500 ng when sperm DNA was not enough) was treated with sodium bisulfite using the EpiTect Bisulfite Kit (QIAGEN) according to the protocol recommended by the manufacturer.
Briefly, 500 ng of genomic DNA was treated with sodium bisulfite using the EpiTect Bisulfite Kit (Qiagen) according to the manufacturer's instructions.
At least 200 ng of DNA was then treated with bisulfite using the EpiTect Bisulfite kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions.
DNAs from BL cell lines or formalin-fixed, paraffin-embedded primary BL cases were modified with bisulfite using the Epitect Bisulfite Kit (Qiagen) according to the manufacturer's protocol, and amplified using primers that did not contain CpG.
The whole volume of amplified negative controls were then mixed with 200 μl Buffer AL and 200 μl ethanol (99%) and purified with the DNeasy Blood and Tissue kit (Qiagen). 1 μg DNA from both individual samples and controls were bisulfite treated using the EpiTect Bisulfite Kit (Qiagen).
600 ng DNA were subsequently bisulfite converted using the EpiTect Bisulfite Kit (Qiagen, Hilden, Germany).
Extracted DNA was treated and modified with a sodium bisulfite procedure using the EpiTect Bisulfite Kit (Qiagen), according to manufacturer's instructions.
The DNA was subsequently resuspended in 500 µL of LoTE (EDTA 2.5 mmol/L and Tris-HCl 10 mmol/L) and stored at −80°C until use. 2 ug of DNA from 28 NSCLCs and 11 normal lung tissues were subjected to bisulfite treatment using the EpiTect® Bisulfite Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions.
