DMMC Course: TECHNIQUES & STRATEGIES IN MOLECULAR MEDICINE

0930-1015 Monday 10 December 2007, Panoz institute, LTEE2, TCD

RNA Detection and quantitation
Dr Shane Duggan (Institute of Molecular Medicine, TCD)

The protein components of the cell are derived by numerous processes indirectly interpreted from a genetic element known as the “gene” which is coded in the cellular DNA. This element is interpreted by the cell in a process called “transcription” where the genetic code for a particular gene is converted into a molecular code known as messenger RNA (mRNA). This mRNA molecule can now be utilised in the creation of a new protein via the translation process. In this lecture the nature and analysis of Ribonucleic acid (RNA) in biological systems will be explored. The understanding of this has allowed the laboratory scientist to interrogate and explore gene expression as it may relate to diseases or cell signalling. Extraction and quantitation of good quality RNA will be discussed as they are the first step in any investigation of gene expression. Standard techniques in common use such as Northern blotting and cycle limited RT-PCR shall also be described as well as more modern techniques such as real time RT-PCR analysis. This lecture will allow the interpretation of published literature utilising these techniques and introduce the steps involved in performing RNA related techniques in your laboratory.