OBJECTIVES: To evaluate the stability of RNA and microRNA (miRNA) in PAXgene-fixed paraffin-embedded tissue blocks after 7 years' storage. METHODS: RNA and miRNA were extracted from PAXgene-fixed paraffin-embedded (PFPE) blocks in 2009 then stored at -80°C. Seven years later, RNA and miRNA were again extracted from the same blocks. RNA and miRNA integrity in the 2009 and 2016 extractions were compared using RNA integrity number (RIN), paraffin-embedded RNA metric (PERM), reverse transcription polymerase chain reaction (RT-PCR) for different amplicon lengths, and quantitative RT-PCR (qRT-PCR) for three mRNA and three miRNA targets. RESULTS: In PFPE blocks, mRNA was poorer in 2016 extractions compared to the 2009 extractions in all blocks and all assays applied, with transcripts degrading at different rates in the same blocks. For miRNA, qRT-PCR showed no statistically significant differences between 2009 and 2016 extractions. CONCLUSIONS: mRNA in PFPE tissue blocks degrades at room temperature storage over 7 years.
OBJECTIVES: To evaluate the stability of RNA and microRNA (miRNA) in PAXgene-fixed paraffin-embedded tissue blocks after 7 years' storage. METHODS: RNA and miRNA were extracted from PAXgene-fixed paraffin-embedded (PFPE) blocks in 2009 then stored at -80°C. Seven years later, RNA and miRNA were again extracted from the same blocks. RNA and miRNA integrity in the 2009 and 2016 extractions were compared using RNA integrity number (RIN), paraffin-embedded RNA metric (PERM), reverse transcription polymerase chain reaction (RT-PCR) for different amplicon lengths, and quantitative RT-PCR (qRT-PCR) for three mRNA and three miRNA targets. RESULTS: In PFPE blocks, mRNA was poorer in 2016 extractions compared to the 2009 extractions in all blocks and all assays applied, with transcripts degrading at different rates in the same blocks. For miRNA, qRT-PCR showed no statistically significant differences between 2009 and 2016 extractions. CONCLUSIONS: mRNA in PFPE tissue blocks degrades at room temperature storage over 7 years.
Authors: Avi Z Rosenberg; Carrie Wright; Karen Fox-Talbot; Anandita Rajpurohit; Courtney Williams; Corey Porter; Olga Kovbasnjuk; Matthew N McCall; Joo Heon Shin; Marc K Halushka Journal: Sci Rep Date: 2018-06-28 Impact factor: 4.379
Authors: Eros Azzalini; Eleonora De Martino; Paolo Fattorini; Vincenzo Canzonieri; Giorgio Stanta; Serena Bonin Journal: Int J Mol Sci Date: 2019-09-27 Impact factor: 5.923
Authors: Mark Z Kos; Sobha Puppala; Dianne Cruz; Jennifer L Neary; Ashish Kumar; Emma Dalan; Cun Li; Peter Nathanielsz; Melanie A Carless Journal: Front Mol Neurosci Date: 2022-03-22 Impact factor: 5.639