GSM4455687: Bisulf_seq_downstream_CTCF_GM04648; Homo sapiens; Bisulfite-Seq
Bisulfite sequencing was performed according to a previously described method (Barbé et al., 2017). Bisulfite conversion was performed with the EZ DNA Methylation kit (Zymo Research) using the standard protocol. 200 ng of genomic DNA were used for bisulfite conversion at 50ºC for 12 hours. Bisulfite-converted DNA was desulphonated, eluted, and immediately used for nested and hemi-nested PCR amplification of the upstream and downstream CTCF binding sites respectively using previously described primers (Barbé et al., 2017). Libraries were constructed by using 50 ng of bisulfite-converted DNA for the first PCR, and 3 µL of the products were used for the second PCR. The final amplicons contained sequencing adapters and were purified with the NucleoSpin PCR Clean-up kit (Macherey-Nagel) and used for 2 x 250 bp paired-end sequencing on Illumina Miseq.
GEO Accession
GSM4455687