A two-step method for the extraction of high-quality RNA from endoscopic biopsies*

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2007 (EN)
A two-step method for the extraction of high-quality RNA from endoscopic biopsies* (EN)

U. Lendeckel, P. Malfertheiner, Th. Wex, G. Treiber,

SUMMARY Background: The usage of molecular techniques such as quantitative RT-PCR depend on the quality of cellular RNA. In particular, RNA extraction from endoscopic biopsies is difficult with respect to yield and, especially, integrity. Therefore, we developed a method that allows extraction of high-quality RNA from these sources. Methods: Endoscopic biopsies taken from the gastric antrum, corpus and duodenum were subjected to various RNA extraction protocols, and the RNA was used for quantitative RT-PCR. Results: The subsequent usage of two methods, (i) a phenol/ chloroform extraction and (ii) a colunn-based extraction method resulted in a yield of 4.5ìg total RNA/biopsy of reliable quality in 80% of samples. The quantitative RTPCR analysis revealed that only RNA samples that clearly show both 18S- and 28S-RNA bands in agarose gel electrophoresis were suitable for quantitative RT-PCR. In all these samples, both the corpus-specific pepsinogen C-mRNA and the duodenum-specific mdr-1-mRNA could be consistently detected. In degraded RNA, pepsinogen C, mdr-1 or â-actin mRNAs were still detectable, but the quantitative det4rmination gave inconsistent data. Conclusions: The two-step method described here proved to be the most reproducible approach to obtaining sufficient amounts of high-quality RNA from endoscopic biopsies as required for quantitative gene expression analyses. (EN)




Hellenic Society of Gastroenterology (EN)

Annals of Gastroenterology; Volume 15, No 2 (2002) (EN)

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