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Selective genetic analysis of p53 immunostain positive cells

Lookup NU author(s): Dr Bridget Wilkins

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Abstract

The isolation of p53 immunostain positive cells from histological sections for molecular genetic studies is a difficult task, especially if there are few positive cells. To eliminate contaminating DNA from p53 negative cells, which can obscure the results of molecular assays, a variation on the technique of immunohistoselective sequencing was developed. This is a highly selective approach, whereby immunostained sections of formalin fixed, paraffin wax embedded tissue are exposed to ultraviolet irradiation to damage the DNA in p53 negative cells. The DNA in positive cells remains unaffected because the dark immunostain protects their nuclei from ultraviolet light. Polymerase chain reaction single strand conformation polymorphism of samples enriched with p53 immunostain positive cells has shown that this method can produce pure samples of mutated DNA. The isolation of DNA from minority immunostain positive cells allows a wide range of molecular analyses to be carried out on these samples, which would otherwise be hampered by the problem of contaminating background cells.

Publication metadata

Author(s): Phelps M, Wilkins BS, Jones DB

Publication type: Article

Publication status: Published

Journal: Molecular Pathology

Year: 2000

Volume: 53

Issue: 3

Pages: 159-61

ISSN (print): 1366-8714

ISSN (electronic): 1472-4154

Publisher: BMJ Group

URL: http://dx.doi.org/10.1136/mp.53.3.159

DOI: 10.1136/mp.53.3.159

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