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Recommendations for mRNA analysis of micro-dissected glomerular tufts from paraffin-embedded human kidney biopsy samples.

Clemens L BockmeyerJuliane WittigKaren SäuberlichPhilipp SelhausenMarc EßerPhilip ZeuschnerFriedrich ModdeKerstin AmannChristoph Daniel
Published in: BMC molecular biology (2018)
Our approach implements compartment-specific glomerular mRNA expression analysis into research applications, even regarding glomerular substructures like parietal epithelial cells. We recommend using of at least 60 micro-dissected unstained glomerular or 300 hemalaun-stained Bowman's capsule transections to obtain sufficient input mRNA for reproducible results. Hereby, the range of RNA concentrations in 60 micro-dissected glomeruli is low and appropriate normalization of Cq values using our suggested reference transcripts (PGK1 and PPIA) allows compensation with respect to different amounts of RNA purity and quantity.
Keyphrases
  • high glucose
  • diabetic nephropathy
  • endothelial cells
  • binding protein
  • working memory
  • nucleic acid
  • ultrasound guided