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

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Annals of Gastroenterology
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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)


Ελληνική Γαστροεντερολογική Εταιρία (EL)
Hellenic Society of Gastroenterology (EN)

Αγγλική γλώσσα


Hellenic Society of Gastroenterology (EN)

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

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