Quantitative DNA Methylation Analysis

Total DNA was extracted from LCLs using the QIAamp^® DNA Mini kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. Two independent quantitative methylation experiments of IGF2, H19, GNAS, GNAS-AS1, MEST and PEG10 DMRs were performed by pyrosequencing using the Pyro Mark ID instrument (Qiagen, Hilden, Germany). Raw data were analyzed using the Q-CpG software v1.09 (Biotage Sweden AB). Details on the genomic positions, set-up and protocol were previously described [45 (link),46 (link)].

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Pileggi S., La Vecchia M., Colombo E.A., Fontana L., Colapietro P., Rovina D., Morotti A., Tabano S., Porta G., Alcalay M., Gervasini C., Miozzo M, & Sirchia S.M. (2021). Cohesin Mutations Induce Chromatin Conformation Perturbation of the H19/IGF2 Imprinted Region and Gene Expression Dysregulation in Cornelia de Lange Syndrome Cell Lines. Biomolecules, 11(11), 1622.

Publication 2021

QIAamp® DNA Mini kit Pyro Mark ID instrument Q-CpG software v1.09

Genomic Igf2 Lcls Methylation

Corresponding Organization : Ospedale San Paolo

Other organizations : Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, University of Insubria, European Institute of Oncology

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Methylation levels of IGF2, H19, GNAS, GNAS-AS1, MEST, and PEG10 differentially methylated regions (DMRs)

control variables

None explicitly mentioned

controls

No positive or negative controls were explicitly mentioned in the input.

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