Streamlined approach to functional analysis of promoter-region polymorphisms

Coleman, Sharon Louise, Hoogendoorn, Bastiaan ORCID: https://orcid.org/0000-0001-9753-169X, Guy, Carol, Smith, Susan Kaye, O'Donovan, Michael Conlon ORCID: https://orcid.org/0000-0001-7073-2379 and Buckland, Paul Robert 2002. Streamlined approach to functional analysis of promoter-region polymorphisms. Biotechniques 33 (2) , pp. 412-418.

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Abstract

We have developed a rapid method for identifying functional promoter-region polymorphisms. Using a modified pGL3 luciferase expression T-vector, we can amplify by PCR, clone, identify allelic pairs of a polymorphic gene promoter region, and prepare plasmids for cell culture 10 promoters (20 allele pairs) per week per researcher. By utilizing 96-well plate technology and an internal control plasmid expressing secreted alkaline phosphatase, each of these allele pairs can be tested for relative promoter activity in each of three cell lines (HEK293t, TE671, and JEG3) with similar resources.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine MRC Centre for Neuropsychiatric Genetics and Genomics (CNGG)
Subjects:	R Medicine > R Medicine (General)
Publisher:	Biotechniques
ISSN:	0736-6205
Last Modified:	12 Dec 2022 08:51
URI:	https://orca.cardiff.ac.uk/id/eprint/57782

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