GSM1824833
GSM1824833: h1ESC dorsomorphin induced 48h_BS-seq; Homo sapiens; Bisulfite-Seq
Qiagen DNeasy mini kit, performed as per manufacturer's instructions For end repair and add dATP, add 7.25 ml EA mix1 and 1.75 ml EA mix 2 to the 16 ml PCR product. Then PCR with the system: 12℃ 15 min; 37℃ 15 min; 72℃ 20 min, and hold in 4℃. Add 2 ml water, 1 ml 10X DNA Ligase Buffer, 2 ml methylated adaptor(5' P-GATCGGAAGAGCTCGTATGCCGTCTTCTGCTTG, 5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT), 5ml T4 DNA ligase to the system, hold overnight in 16℃ for 16 hours. Add Agencourt® AMPure® XP to purify the system with the manufature-provided manual, and get 200 ml product. Add 6 ml Loading Dye(6X), vertex and perform  2% agarose gel electrophoresis with 90V for 2 hours, select 300bp-400bp fragments for gel extraction with Qiagen Minilute Gel Purification Kit by following its protocol. Solute the product in 20 ml EB buffer. MethylCode™ Bisulfite Conversion Kit for bisulfite conversion was used by following protocol for the kit. Purify the product with EZ DNA Methylation-Gold Kit and get 10 ml product. Add 1 ml Bioo Primer, 1 ml water and 12.5 ml KAPA HiFi Hotstart Utracil+ ReadyMix (2X) to the system. PCR: 95℃ 10mins, 5 cycles: 95℃ 30s, 65℃ 30s, 72℃ 45s, 72℃ 7min, and hold in 4℃. Then purify with Agencourt® AMPure® X Beads.
GEO
GSM1824833
GEO Accession
GSM1824833