Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

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Evaluation of the efficacy of various reagents in improving microRNA extraction.

Author(s): Kikuchi A, Naruse A, Sawamura T, Nonaka K

Publication: Ann Clin Biochem, 2019, Vol. 56, Page 375-380

PubMed ID: 30813744 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine if miRNA extraction efficiency using the HighPure miRNA isolation kit could be improved through the use of an alternative binding enhancer. Ten surgically-resected FFPE colon cancer specimens were obtained (additional details not provided). miRNA was extracted from xylene deparaffinized 5 µm sections using the HighPure miRNA isolation kit in conjunction with the optimal volume (experimentally determined) of different binding enhancer solutions. The tested binding solutions included 100 µL of the Roche binding enhancer solution (BE), 400 µL of 2-butanol, 400 µL of 2-propanol, 300 µL of acetone, 300 µL of acetonitrile, 400 µL of ethanol, 200 µL of PEG 1540, 200 µL of PEG 2000, 300 µL of tetraethylene glycol dimethyl ether (TDE), 300 µL of tetrahydrofuran (THF), and 500 µL of 1,4-dioxane. miR-21 levels following extraction were determined by real-time RT-PCR and compared to that using the Roche binding enhancer solution.

   Summary of Findings:

   The mean cycle threshold values for miR-21 were 1.91-2.83 cycles lower using the alternative binding solutions than when the Roche binding enhancer solution was used, indicating an increased efficiency of extraction. The largest decrease in mean cycle threshold values were obtained using PEG 1540 and PEG 2000 (2.42 and 2.83, respectively). PEG was seen as particularly advantageous as the optimal volume was lower than for the other solutions tested.

   Biospecimens

   • Tissue - Colorectal

   Preservative Types

   • Formalin

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Real-time qRT-PCR Specific	Targeted nucleic acid	miR-21
   Analyte Extraction and Purification	Analyte isolation method	HighPure miRNA isolation kit with the Roche binding enhancer solution HighPure miRNA isolation kit with candidate binding solution HighPure miRNA isolation kit with 2-propanol HighPure miRNA isolation kit with ethanol HighPure miRNA isolation kit with PEG 600 HighPure miRNA isolation kit with PEG 1000 HighPure miRNA isolation kit with PEG 1540 HighPure miRNA isolation kit with PEG 2000 HighPure miRNA isolation kit with TDE HighPure miRNA isolation kit with THF HighPure miRNA isolation kit with 1,4-dioxane HighPure miRNA isolation kit with 2-butanol

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