Sonification as a means of enhancing the detection of gene expression levels from formalin-fixed, paraffin-embedded biopsies
Houze, T.A.; Gustavsson, B.
Biotechniques 21(6): 1074-8 1080 1082
1996
ISSN/ISBN: 0736-6205 PMID: 8969836 Document Number: 462070
We describe a novel approach for the isolation and quantification of mRNA from archival specimens by using oligo(dT)-25 paramagnetic beads and the reverse transcription polymerase chain reaction (RT-PCR). A preliminary study is conducted to evaluate experimental conditions and the effects of varied periods of sonification on the yield of mRNA isolated from formalin-fixed, paraffin-embedded human biopsy samples. Validation of this approach will involve the comparison of observed beta-actin gene expression levels among different samples from the same patient after various periods of sonification. The novelty of this method is that it offers a means of analyzing archival samples using semiquantitative RT-PCR following sonification-enhanced mRNA extraction. The utility of this protocol allows for gene expression studies to be done effectively using the formalin-fixed, paraffin-embedded archival samples that are commonly available in all hospital pathology departments.