English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Mutation screening in 86 known X-linked mental retardation genes by droplet-based multiplex PCR and massive parallel sequencing

Hu, H., Wrogemann, K., Kalscheuer, V., Tzschach, A., Richard, H., Haas, S. A., et al. (2009). Mutation screening in 86 known X-linked mental retardation genes by droplet-based multiplex PCR and massive parallel sequencing. Hugo J, 3(1-4), 41-9. doi:10.1007/s11568-010-9137-y.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/21.11116/0000-0002-E02B-1 Version Permalink: http://hdl.handle.net/21.11116/0000-0002-E02C-0
Genre: Journal Article

Files

show Files

Locators

show
hide
Description:
-

Creators

show
hide
 Creators:
Hu, H.1, Author
Wrogemann, K.1, Author
Kalscheuer, V.1, Author
Tzschach, A.1, Author
Richard, H.1, Author
Haas, S. A.1, Author
Menzel, C.1, Author
Bienek, M.1, Author
Froyen, G.1, Author
Raynaud, M.1, Author
Van Bokhoven, H.1, Author
Chelly, J.1, Author
Ropers, H.1, Author
Chen, W.1, Author
Affiliations:
1Max Planck Society, ou_persistent13              

Content

show
hide
Free keywords: -
 Abstract: UNLABELLED: Massive parallel sequencing has revolutionized the search for pathogenic variants in the human genome, but for routine diagnosis, re-sequencing of the complete human genome in a large cohort of patients is still far too expensive. Recently, novel genome partitioning methods have been developed that allow to target re-sequencing to specific genomic compartments, but practical experience with these methods is still limited. In this study, we have combined a novel droplet-based multiplex PCR method and next generation sequencing to screen patients with X-linked mental retardation (XLMR) for mutations in 86 previously identified XLMR genes. In total, affected males from 24 large XLMR families were analyzed, including three in whom the mutations were already known. Amplicons corresponding to functionally relevant regions of these genes were sequenced on an Illumina/Solexa Genome Analyzer II platform. Highly specific and uniform enrichment was achieved: on average, 67.9% unambiguously mapped reads were derived from amplicons, and for 88.5% of the targeted bases, the sequencing depth was sufficient to reliably detect variations. Potentially disease-causing sequence variants were identified in 10 out of 24 patients, including the three mutations that were already known, and all of these could be confirmed by Sanger sequencing. The robust performance of this approach demonstrates the general utility of droplet-based multiplex PCR for parallel mutation screening in hundreds of genes, which is a prerequisite for the diagnosis of mental retardation and other disorders that may be due to defects of a wide variety of genes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11568-010-9137-y) contains supplementary material, which is available to authorized users.

Details

show
hide
Language(s):
 Dates: 2009
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: Other: 21836662
DOI: 10.1007/s11568-010-9137-y
ISSN: 1877-6566 (Electronic) 1877-6558 (Linking)
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Hugo J
  Alternative Title : The HUGO journal
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 3 (1-4) Sequence Number: - Start / End Page: 41 - 9 Identifier: -