14/03/05 00:19:58.53
Bisulphite sequencing
URLﾘﾝｸ(www.ncbi.nlm.nih.gov)
Bisulfite treatment of DNA was done using
the CpGenome DNA Modification Kit (Chemicon, Temecula, CA,
URLﾘﾝｸ(www.chemicon.com)<)), following the manufacturer’s
instructions. The resulting modified DNA was amplified by nested PCR using two forward (F)
primers and one reverse (R) primer: Oct4 (F1, 5′-GTTGTTTTGTTTTGGTTTTGGATAT-3′; F2,
5′-ATGGGTTGAAATATTGGGTTTATTTA-3′; R, 5′-CCACCCTCTAACCTTAACCTCTAAC-3′). And
Nanog (F1, 5′-GAGGATGTTTTTTAAGTTTTTTTT-3′; F2, 5′-AATGTTTATGGTGGATTTTGTAGGT-3′; R,
5′-CCCACACTCATATCAATATAATAAC-3′).