GSM1873499: NIH3T3_RRBS_EP10; Mus musculus; Bisulfite-Seq
Genomic DNA were extracted using Easy-DNA™ gDNA Purification Kit (Life Technologies). Total RNA were extracted using TRIZol reagent. For RRBS, 1 μg of genomic DNA from stable clones overexpressing 14-3-3ε or control vector was digested by MspI, end-repaired and dA-tailed according to manufacturer’s conditions (New England Biolabs). Methylated NEB Illumina loop adaptor was ligated to digested DNA (E7370S, New England Biolabs). Ligation product was size-selected for 150 to 400 bp fragments on 2% agarose gels and bisulfite converted using the EZ DNA Methylation Kit (Zymo Research). Libraries were enriched by PCR using EpiMark Hot Start Taq DNA Polymerase (New England Biolabs). For RNA-seq, libraries were constructed using the NEBNext® Ultra™ Directional RNA Library Prep Kit (New England Biolabs) per the manufacturer’s instructions. Average insert size of libraries was 200 bp.
GEO Accession
GSM1873499