Anna Lehmann's present address is South West Thames Regional Genetics Service, St George's, University of London, UK
Targeted methylation testing of a patient cohort broadens the epigenetic and clinical description of imprinting disorders
Article first published online: 2 AUG 2013
Copyright © 2013 Wiley Periodicals, Inc.
American Journal of Medical Genetics Part A
Volume 161, Issue 9, pages 2174–2182, September 2013
How to Cite
2013. Targeted methylation testing of a patient cohort broadens the epigenetic and clinical description of imprinting disorders. Am J Med Genet Part A 161A:2174–2182., , , , , , , , , , .
Conflict of interests: None.
Consortium members are listed as an Appendix in the Supplementary Information.
- Issue published online: 14 AUG 2013
- Article first published online: 2 AUG 2013
- Manuscript Accepted: 13 MAR 2013
- Manuscript Received: 8 NOV 2012
- Newlife Foundation for Disabled Children
- Hampshire and the Isle of Wight NIHR Comprehensive Local Research Network
Additional supporting information may be found in the online version of this article at the publisher's web-site.
|ajmga36049-sm-0001-SupMat-S2.doc||34K||Supplementary Material 2: A table detailing the clinical features of individuals with SRS-HIL|
|ajmga36049-sm-0001-SupMat-S3.doc||38K||Supplementary Material 3: A table detailing the clinical features of individuals with BWS-HIL|
|ajmga36049-sm-0001-SupFig-S1.tif||1521K||Figure S1. Molecular analysis of ICR2 in patients with clinical features of SRS Panel A: schematic representation of KCNQ1 and the imprinting control region ICR2 in the KCNQ1OT1 promoter. Top panel: Location of KCNQ1OT1 (blue line) and maternally-methylated ICR2 (red lollipops) within KCNQ1 (red dashed line with exons indicated as vertical bars). Scale (in Mb) is according to HG18. Second panel: ICR 2 region showing location of amplicons for molecular analysis. The transcription origin of KCNQ1OT1 is represented by a blue arrow, ICR2 by a red bar. Locations of amplicons for genomic sequencing, MS-PCR, bisulphite sequencing and pyrosequencing are indicated by black bars. Panel B: Pyrograms of ICR2 amplicon 4 in patients and control. Each panel represents a representative pyrogram of the first bisulphite-induced C/T polymorphism in pyrosequencing reaction D. The percentage cytosine (i.e., percentage methylation) is marked over each pyrogram. Panel C: Pyrosequencing analysis of ICR2 in the three patients and a normal control. For each patient the methylation index is represented as the percentage cytosine at the first bisulphite-induced polymorphism in the pyrosequence, as an average of duplicate tests. The control value is the mean and standard deviation of duplicate determinations of four normal controls at the same position. Panel D: Summary of methylation index as determined by methylation-specific MLPA. For each patient the methylation index is stated for each of the four probes within ICR2, and the mean and standard deviation of the five probes within ICR1. In a normal individual the methylation indices at both ICR2 and ICR1 would be expected to approximate to 1.0, with abnormal calls being made outside the range 0.75-1.3. Panel E: Bisulphite sequence analysis of ICR2 in the three patients, a normal control, and a control with BWS caused by ICR2 hypomethylation. Filled and empty circles represent methylated and unmethylated cytosines, respectively, within CG dinucleotides in the amplicon. Numbers to the right of each sequence represent the number of clones, out of a total of 20, in which that sequence was present.|
|ajmga36049-sm-0001-SupMat-S5.doc||56K||Supplementary Material 5: Clinical features of individuals referred for SRS diagnostic testing, with an unsuspected anomaly of imprinting.|
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