GSM6398669: CD1-brain-M-5M-DNA-5; Mus musculus; OTHER
Dounce homogenizers were used for each cortex sample in a general lysis buffer containing 0.1 M Tris-pH8, 0.2M NaCl, 5 mM EDTA, 10% SDS, 150 mM DTT and proteinase K. The genomic DNA was extracted from the lysate using phenol:chloroform:isoamyl alcohol extraction followed by EtOH precipitation. 2 µg of genomic DNA was sheared with Diagenode Bioruptor to yield DNA fragments that are approximately 300 bp. DNA fragments were end repaired, A-tailed, and ligated to Illumina adaptors followed by PCR amplification for 7 cycles. The final libraries were size selected using AMPure XP beads, so the fragments were between the size of 200-400 bp long. Each library was subject to paired-end sequencing for 150 bp reads using Illumina NovaSeq platform.