GSM6588822: colon, LCM, G205, NEC; Homo sapiens; Bisulfite-Seq
Snap-frozen specimens were mounted on appropriate embedding molds with clear OCT compound and sectioned on a cryostat instrument. These sections were mounted on membrane slides, stained with toluidine blue and air-dried. Laser dissection and capture of the chosen tissue region of interest (ROI) were cut by the laser capture microdissection (LCM) Instrument. The ROI laser capture tissue samples were subsequently collected into appropriate PCR tube for further DNA extraction and subsequent studies. DNA was extracted from LCM tissue using the Nucleospin Tissue XS Kit (Macherey–Nagel) with the modifcations suggested for LCM. DNA was sheared with Covaris to a size of ~175bp. Libraries were prepared using the KAPA HyperPrep Kit. Libraries were bisulfite converted post-ligation using the EZ DNA Methylation-Direct Kit and amplified 12 cycles. Libraries were sequenced on an Illumina HiSeqX using 150-bp paired-end reads.