Abstract：
:We have developed a high-throughput, multiplex reverse transcription PCR (RTPCR) assay that is suitable for the analysis of medium-to low-copy cellular RNA transcripts from small numbers of cells (10(4)). High throughput was attained by utilizing microplate-based RNA extraction and RTPCR protocols, followed by PCR product visualization of a multiwelled agarose gel, stained with SYBR Green I dye. The transcriptional assay was unaffected by solvents (dimethyl sulfoxide and methanol) routinely used in high-throughput drug screens at concentrations required for compound solubilization. Furthermore, it has been used successfully for the investigation of differential mRNA expression levels of tumor necrosis factor alpha (TNF-alpha) and Interleukin-1 beta (IL-1 beta) in lipopolysaccharide (LPS)-stimulated THP-1 cells (a human monocytic cell line) and the identification of specific IL-1 beta transcriptional inhibitors.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Su S,Vivier RG,Dickson MC,Thomas N,Kendrick MK,Williamson NM,Anson JG,Houston JG,Craig FFdoi
10.2144/97226st02subject
Has Abstractpub_date
1997-06-01 00:00:00pages
1107-13issue
6eissn
0736-6205issn
1940-9818journal_volume
22pub_type
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