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.