Abstract：
:RNA isolated from fixed and paraffin-embedded tissues is widely used in biomedical research and molecular pathology for diagnosis. In the present study, we have set-up a method based on high performance liquid chromatography (HPLC) to investigate the effects of different fixatives on RNA. By the application of the presented method, which is based on the Nuclease S1 enzymatic digestion of RNA extracts followed by a HPLC analysis, it is possible to quantify the unmodified nucleotide monophosphates (NMPs) in the mixture and recognize their hydroxymethyl derivatives as well as other un-canonical RNA moieties. The results obtained from a set of mouse livers fixed/embedded with different protocols as well from a set of clinical samples aged 0 to 30 years-old show that alcohol-based fixatives do not induce chemical modification of the nucleic acid under ISO standard recommendations and confirm that pre-analytical conditions play a major role in RNA preservation.
journal_name
Int J Mol Scijournal_title
International journal of molecular sciencesauthors
Fattorini P,Forzato C,Tierno D,De Martino E,Azzalini E,Canzonieri V,Stanta G,Bonin Sdoi
10.3390/ijms21207540subject
Has Abstractpub_date
2020-10-13 00:00:00issue
20issn
1422-0067pii
ijms21207540journal_volume
21pub_type
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