GSM1838400: RRBseq_Smad1_5_FL_2; Mus musculus; Bisulfite-Seq
gDNA extraction was performed using the Wizard Genomic DNA purification kit (Promega) gDNA concentration was measured with Quant-it Picogreen dsDNA assay and 1 μg input was used in the MSP1 digest. Following overnight incubation at 37°C, digestion reactions were terminated by adding 0.5 M EDTA and purified on a GeneJET PCR purification column. Libraries were prepped using the NEBNext Ultra DNA library preparation kit for Illumina and methylated adapters (Index primers Set 1). Subsequently, adapter ligated fragments were BS converted using the EZ DNA Methylation Gold kit (Zymo Research). 14 cycles of PCR were performed and the products were purified using AMPure XP beads. Quality of the final libraries was checked on a High sensitivity DNA chip (Agilent) and concentration was measured with qPCR. Sequencing was done on an Illumina HiSeq2500 PE 2x50bp
GEO Accession
GSM1838400