I'm trying to evaluate for mRNA expression level of ONE gene in hundreds of tumor samples. What is the method to do this?
Edit: Also, does the type of tissue I have affect which method to use? For example, frozen tissue vs FFPE tissue.
Hi,
The best method used to evaluate mRNA expression is Quantitative RT-PCR in order to quantitate the levels of mRNA species and to detect mRNA in small numbers of cells.
Hope this may help you.
Thank you for responding. Is RT-PCR the most efficient way to evaluate mRNA expression in hundreds of tumor samples? Or are there high throughput assays that can evaluate this better? I only need the expression level of one gene though, so not sure if some of the microarrays or other assays would be useful.
Hi Lingjuan Wang-Li,
If you are able to obtain good yield of RNA after isolating from formalin-fixed, paraffin-embedded tissue sections OR cryo preserved tissues both in terms of quantity and quality, you may opt for RT-PCR based assay. Although it might be labor-intensive for performing this assay in hundreds of samples but this method is certainly more economic, reproducible, robust and free from inter-individual and intra-individual assay variations.
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