Streamlined Approach to Functional Analysis of Promoter-Region Polymorphisms | Zendy

Sharon L. Coleman | Zendy; Bastiaan Hoogendoorn | Zendy; C Guy | Zendy; S. K. Smith | Zendy; Michael O’Donovan | Zendy; Paul R. Buckland | Zendy

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Streamlined Approach to Functional Analysis of Promoter-Region Polymorphisms

Author(s) -

Sharon L. Coleman,

Bastiaan Hoogendoorn,

C Guy,

S. K. Smith,

Michael O’Donovan,

Paul R. Buckland

Publication year - 2002

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/02332ht01

Subject(s) - promoter , allele , plasmid , genetics , luciferase , biology , gene , clone (java method) , promoter activity , microbiology and biotechnology , hek 293 cells , transfection , gene expression

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.

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