Design of a real time quantitative PCR assay to assess global mRNA amplification of small size specimens for microarray hybridisation.

ABSTRACT:

Background

Low RNA yields from clinical samples are a limiting step for microarray technology.

Aims

To design an accurate real time quantitative polymerase chain reaction (PCR) assay to assess the crucial step of global mRNA amplification performed before microarray hybridisation, using less than 1 microg total RNA.

Methods

Three RNA extraction procedures were compared for small size samples. Total RNA was amplified from universal RNA or the BC-H1 breast cancer micrometastatic cell line using three different protocols. Real time quantitative PCR technology was used for accurate measurement of urokinase plasminogen activator receptor and cytokeratin 8 RNA amplification rates and ratios, using primer sets binding at various distances from the 3' end of transcripts. A 50

SUBMITTER: Choesmel V 

PROVIDER: S-EPMC1770498 | biostudies-literature | 2004 Dec

REPOSITORIES: biostudies-literature