GSM8487390: RRBS_Apc_Kras_G13D_tumor_Rep2; Mus musculus; Bisulfite-Seq
Tumors were isolated during necropsy of moribund animals and immediately snap frozen in liquid nitrogen. The animal procedures were performed under protocol approved by the Beth Israel Deaconess Medical Center IACUC. Concentration of genomic DNA (gDNA) was quantified using a Qubit 3.0 Fluorometer. RRBS was performed on 100 ng gDNA of each sample using the NuGen Ovation RRBS Methyl-Seq System (Tecan, #0353) following the manufacturer's recommendationswith the following modifications:After the final repair step, thebisulfite conversion of DNA was conducted using theQiagen EpiTect Fast Bisulfite Conversion kit(Qiagen, #59824) following the manufacturer's recommendations, eluting the bisulfite converted DNA in 23 µl EB. Libraries were amplified with 12 cycles of PCR. Amplified library purificationwith Agencourt RNAclean XP beads (Beckman Coulter, #A63987)was performed twice (1X). The purified libraries were sequencedfor 76 bp single-end readson a NovaSeq 6000 platform (Illumina).