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Mol Biotechnol. 2014 Apr;56(4):312-8. doi: 10.1007/s12033-013-9710-4.

Perspective on the technical challenges involved in the implementation of array-CGH in prenatal diagnostic testing.

Author information

1
Wessex Regional Genetics Laboratory, Salisbury District Hospital, Salisbury, Wiltshire, SP2 8BJ, UK, jonathan.callaway@salisbury.nhs.uk.

Abstract

Our aim was to construct a streamlined technical workflow to facilitate a prospective, multi-centre evaluation of array comparative genomic hybridisation (array-CGH) in the prenatal diagnostic context. A collection of commercially available DNA extraction and quantification techniques were evaluated and compared using minimal quantities of amniotic fluid, chorionic villi and cultured cells. When prenatal DNA of suitable quality and quantity was obtained, array-CGH was performed using Oxford Gene Technology's (OGT, Oxford, UK) CytoSure™ ISCA 8 × 60 K oligo array platform. With starting quantities of 2-4 ml amniotic fluid, 2-5 mg chorionic villi or under 150,000 cultured cells the following optimised technical workflow was identified: DNA extraction using the iGENatal™ kit (igenbiotech, Madrid, Spain) and quantification by the Qubit® 2.0 Fluorometer with the Qubit® dsDNA BR assay kit (Invitrogen™, Eugene, OR, USA). In addition, it was elucidated that array-CGH can be successfully performed with as little as 125 ng DNA in the experiment using the OGT CytoSure™ ISCA 8 × 60 K oligo array platform. Amidst an on-going debate on whether array-CGH should be applied in the prenatal diagnostic setting, by following the technical recommendations described here genetics laboratories can now gain exposure to prenatal array-CGH testing without compromising the conventional karyotype result.

PMID:
24146428
DOI:
10.1007/s12033-013-9710-4
[Indexed for MEDLINE]

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