The development of a protocol for the analysis of genetic expression through «differential display», as a means to reducing the number of false positives

Authors

  • JM VIEITES Departamento de Bioquímica y Biología Molecular. Facultad de Farmacia. Universidad de Granada Granada
  • A SÁNCHEZ-POZO Departamento de Bioquímica y Biología Molecular. Facultad de Farmacia. Universidad de Granada Granada
  • A GIL Departamento de Bioquímica y Biología Molecular. Facultad de Farmacia. Universidad de Granada Granada
  • A SUÁREZ Departamento de Bioquímica y Biología Molecular. Facultad de Farmacia. Universidad de Granada Granada

Keywords:

Differentiation, Enterocytes, Gene expression analysis

Abstract

The analysis of genetic expression, the differential display (DD) method has been widely used, but inspite of the extensive use of the «microarrays» method, it is still to be considered as a valid methodfor the analysis of samples whose transcriptone is not known. In this work, an attempt has been madeto reduce the high number of false positives generated by this technique by optimising method protocol.As a preliminary step, we radioactively marked the oligo dT primer with which the fragments ofidentified DNA were extreme 3'-UTR of mRNA. For each sample two inverse transcriptions and twoPCR reactions were performed. Only fragments of DNA that are expressed differentially in all 4 PCRreactions should be selected. Finally, all of the fragments were cloned and sequenced in triplicate. Theseprotocol modifications have allowed us to identify 5 differentially expressed genes, in intestinal epithelialcells in both proliferative and differentiated states.

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Published

2005-03-20

How to Cite

1.
VIEITES J, SÁNCHEZ-POZO A, GIL A, SUÁREZ A. The development of a protocol for the analysis of genetic expression through «differential display», as a means to reducing the number of false positives. Ars Pharm [Internet]. 2005 Mar. 20 [cited 2024 May 21];46(2):193-204. Available from: https://revistaseug.ugr.es/index.php/ars/article/view/5075

Issue

Vol. 46 No. 2 (2005)

Section

Original Articles

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