[HTML][HTML] Quantitative gene expression analysis in renal biopsies: a novel protocol for a high-throughput multicenter application
CD Cohen, K Frach, D Schlöndorff, M Kretzler - Kidney international, 2002 - Elsevier
CD Cohen, K Frach, D Schlöndorff, M Kretzler
Kidney international, 2002•ElsevierQuantitative gene expression analysis in renal biopsies: A novel protocol for a high-
throughput multicenter application. Background Recent advances in gene expression
analysis may add the quantification of mRNA species in renal biopsies to routine diagnostic
procedures in nephrology. Methods A systematic evaluation was performed on the relevant
steps required to efficiently obtain cDNA from renal biopsies for high-throughput reverse
transcription-polymerase chain reaction (RT-PCR) based mRNA quantification. Results The …
throughput multicenter application. Background Recent advances in gene expression
analysis may add the quantification of mRNA species in renal biopsies to routine diagnostic
procedures in nephrology. Methods A systematic evaluation was performed on the relevant
steps required to efficiently obtain cDNA from renal biopsies for high-throughput reverse
transcription-polymerase chain reaction (RT-PCR) based mRNA quantification. Results The …
Quantitative gene expression analysis in renal biopsies: A novel protocol for a high-throughput multicenter application.
Background
Recent advances in gene expression analysis may add the quantification of mRNA species in renal biopsies to routine diagnostic procedures in nephrology.
Methods
A systematic evaluation was performed on the relevant steps required to efficiently obtain cDNA from renal biopsies for high-throughput reverse transcription-polymerase chain reaction (RT-PCR) based mRNA quantification.
Results
The protocol preserves mRNA integrity by a novel RNase inhibitor and allows meticulous microdissection followed by maximal RNA recovery from tissue samples. Reverse transcription was optimized to give the best yield from minimal starting material. RNA quantity and quality were systematically investigated by real-time RT-PCR and electrophoresis on a microfluidic system, respectively. The reported procedure offers high RNA preservation and increases the yield of cDNA significantly compared to former protocols.
Conclusion
The simplicity of biopsy material acquisition combined with the centrally performed processing makes this protocol suitable for a wide spectrum of expression analysis in diverse clinical settings.
Elsevier