GSM3756095: Male_Sult3a2_Repl2_G145_M36; Mus musculus; OTHER
Nuclei were isolated by ultracentrifugation in a high sucrose buffer then crosslinked with 0.8% formaldehyde for 9 minutes. 10M Crosslinked nuclei were processed as described previously (van de Werken et al 2012, Splinter et al 2012). DpnII was used as the primary restriction enzyme and Csp6I was used as the secondary. All ligations and disgestions were overnight (minimum 16 hr), and were verified by running a small aliquot in a 1% agarose gel. Samples were amplified for 25 cycles of inverse PCR using a custom primer pair containing sequence specific to the indicated viewpoint as well as a partial adaptor sequence to facilitate barcoding in a secondary PCR reaction. To reduce the impact of PCR domination, samples represent pools of 6 independent (but otherwise identical) inverse PCR reactions. The reading primer was anchored directly at the DpnII cutsite, while the nonreading primer was positioned ~75bp from the Csp6I site. The samples were barcoded using New England Biolabs NEBNext Multiplex Oligos for Illumina (#E7335) with 5 rounds of additional PCR per the manufacturer's instructions.
GEO Accession
GSM3756095