دورية أكاديمية
A simple procedure for routine RNA extraction and miRNA array analyses from a single thyroid in vivo fine needle aspirate.
| العنوان: | A simple procedure for routine RNA extraction and miRNA array analyses from a single thyroid in vivo fine needle aspirate. |
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| المؤلفون: | Rossing M; Department of Clinical Biochemistry, Rigshospitalet, Copenhagen, Denmark. maria.rossing@rh.regionh.dk, Kaczkowski B, Futoma-Kazmierczak E, Glud M, Klausen M, Faber J, Nygaard B, Kiss K, Sørensen CH, Nielsen FC, Bennedbæk FN, Friis-Hansen L |
| المصدر: | Scandinavian journal of clinical and laboratory investigation [Scand J Clin Lab Invest] 2010 Dec; Vol. 70 (8), pp. 529-34. Date of Electronic Publication: 2010 Oct 15. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Informa Healthcare Country of Publication: England NLM ID: 0404375 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1502-7686 (Electronic) Linking ISSN: 00365513 NLM ISO Abbreviation: Scand J Clin Lab Invest Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: London : Informa Healthcare Original Publication: Oslo, Medisinsk Fysiologisk Forenings Forlag. |
| مواضيع طبية MeSH: | MicroRNAs/*genetics , MicroRNAs/*isolation & purification , Oligonucleotide Array Sequence Analysis/*methods , Thyroid Gland/*metabolism , Thyroid Gland/*pathology, Biopsy, Fine-Needle ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; MicroRNAs/metabolism ; Reproducibility of Results |
| مستخلص: | Context: microRNA (miRNA) expression profiling and classification of tissue obtained from fine-needle aspirates (FNA) could be a major improvement of the preoperative diagnosis of thyroid nodules. Objective: Before this can be clinically implemented, a robust and non-toxic method for obtaining sufficient quantity and quality of RNA from single in vivo FNA has to be established. RNAlater is a non-toxic RNA-stabilizing agent. However, due to the high density of RNAlater, pelleting of the tissue samples is difficult, and results in low recovery of RNA that is insufficient for subsequent miRNA array expression analysis. We therefore developed a simple centrifugation method for capturing tissue stored in RNAlater on a 0.45-μm filter. Design: FNA from 24 patients with a solitary cold thyroid nodule was stored in Trizol, liquid nitrogen, or RNAlater. The tissue stored in RNAlater was either pelleted by centrifugation or captured on the 0.45-μm filters. RNA was extracted using the Trizol method. To validate results, FNA from additional 30 patients were analyzed based on the modified RNAlater protocol. Main Outcome: Capturing FNA tissue samples on the filters increased the RNA yield 10 fold and maintained RNA purity, permitting miRNA array expression profiling and allowing comparable levels of known miRNA-clusters regardless of preservation technique. Results were confirmed in an additional 30 patients. Conclusion: The modified RNAlater protocol is well suited for isolating RNA from single thyroid in vivo FNA in a clinical setting. Furthermore this permits shipping of FNA samples at room temperature from peripheral centers to a centralized array core facility. |
| المشرفين على المادة: | 0 (MicroRNAs) |
| تواريخ الأحداث: | Date Created: 20101019 Date Completed: 20110330 Latest Revision: 20101118 |
| رمز التحديث: | 20240628 |
| DOI: | 10.3109/00365513.2010.522250 |
| PMID: | 20950121 |
| قاعدة البيانات: | MEDLINE |
PDF full text from Publisher | تدمد: | 1502-7686 |
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| DOI: | 10.3109/00365513.2010.522250 |