Optimized RNA isolation of FFPE uterine scar tissues for RNA expression analyses delineated by laser microdissection

Optimized RNA isolation of FFPE uterine scar tissues for RNA expression analyses delineated by laser microdissection

Samples for histological analyses are often formalin-fixed paraffin-embedded (FFPE) and slide-mounted, which complicates RNA extraction for many downstream molecular applications. Furthermore, when the region of interest is extremely small due to isolation with laser microdissection (LMD), extractin...

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Veröffentlicht in:BioTechniques. - 1988. - 72(2022), 6 vom: 24. Juni, Seite 273-278
1. Verfasser: Paping, Alexander (VerfasserIn)
Weitere Verfasser: Basler, Clara, C Rancourt, Rebecca, Ehrlich, Loreen, Melchior, Kerstin, Henrich, Wolfgang, Braun, Thorsten
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2022
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Research Support, Non-U.S. Gov't FFPE LMD RNA expression RNA isolation laser microdissection qRT-PCR uterine scar tissue Formaldehyde mehr... 1HG84L3525 RNA 63231-63-0
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520 |a Samples for histological analyses are often formalin-fixed paraffin-embedded (FFPE) and slide-mounted, which complicates RNA extraction for many downstream molecular applications. Furthermore, when the region of interest is extremely small due to isolation with laser microdissection (LMD), extracting RNA of adequate quality and quantity is difficult. We describe an optimized protocol for maximizing RNA output from FFPE tissue devised to identify and analyze gene expression of human maternal uterine scar tissue obtained from uterotomy scars resulting from prior cesarean deliveries. Gomori trichrome staining allowed for region identification for LMD. Successful RNA isolation, reverse transcription and, importantly, quantitative real-time PCR (qRT-PCR) were performed. This report provides an optimized step-by-step protocol yielding sufficient RNA for qRT-PCR analyses from challenging tissue/LMD-FFPE samples 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
650 4 |a FFPE 
650 4 |a LMD 
650 4 |a RNA expression 
650 4 |a RNA isolation 
650 4 |a laser microdissection 
650 4 |a qRT-PCR 
650 4 |a uterine scar tissue 
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650 7 |a 1HG84L3525  |2 NLM 
650 7 |a RNA  |2 NLM 
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700 1 |a Basler, Clara  |e verfasserin  |4 aut 
700 1 |a C Rancourt, Rebecca  |e verfasserin  |4 aut 
700 1 |a Ehrlich, Loreen  |e verfasserin  |4 aut 
700 1 |a Melchior, Kerstin  |e verfasserin  |4 aut 
700 1 |a Henrich, Wolfgang  |e verfasserin  |4 aut 
700 1 |a Braun, Thorsten  |e verfasserin  |4 aut 
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