Peripheral blood samples were collected from all subjects. About 5 ml of peripheral venous blood was taken from the elbow using an EDTA anticoagulant tube (before surgery for patients with thyroid nodules). The whole blood samples were subjected to immediate centrifugation at 1,000 x g and 4˚C for 10 min. Plasma obtained from the supernatant was dispensed in 300-µl aliquots into cryotubes and stored at -80˚C. Circulating RNAs were extracted through multiple steps including homogenization, phase separation, RNA precipitation, RNA wash and RNA re-dissolving. The final RNA solution was stored at -80˚C. Following quality control tests of the extracted RNA, miRNAs were subsequently labeled with fluorescent dyes. Samples containing labeled miRNAs were then hybridized with miRCURY LNATM microRNA Array (v.18.0, Exiqon). The images were scanned and analyzed. Abnormally expressed miRNAs found through microarray analysis were further validated by real-time quantitative PCR using plasma samples from patient subjects. All tissue samples and postoperative plasma samples were subjected to real-time quantitative PCR in order to verify whether the above abnormally expressed miRNAs were also abnormally expressed in these samples.
Mircury lnatm microrna array
Manufactured by Qiagen
Sourced in Denmark
The MiRCURY LNA™ microRNA Array is a laboratory analysis tool designed for the detection and profiling of microRNA (miRNA) expression. It provides a comprehensive solution for the analysis of miRNA expression patterns in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Agilent feature extraction software version 10.7.3.1, by Agilent Technologies (1 mentions)
Hybridization chamber kit part g2534a, by Agilent Technologies (1 mentions)
Hybridization gasket slide kit part g2534 60003, by Agilent Technologies (1 mentions)
Agilent g2565ca microarray scanner system, by Agilent Technologies (1 mentions)
Pcr master mix, by Arraystar (1 mentions)
Mircury array power labeling kit, by Qiagen (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Array pro analyzer, by Media Cybernetics (1 mentions)
Genepix 4000b, by Molecular Devices (1 mentions)
4 protocols using mircury lnatm microrna array
1
Plasma miRNA Profiling in Thyroid Nodules
2
miRNA Expression Profiling by Microarray
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Human microRNA expression was analyzed with miRCURY LNATM microRNA Array (Exiqon, Vedbaek, Denmark), covering 1918 well-characterized human microRNAs among 3100 capture probes for human, mouse, and rat miRNAs. In this procedure, 5′-phosphate from 250 ng of total RNA was removed by treating with Calf Intestinal Alkaline Phosphatase (CIP) followed by labeling with Hy3 green fluorescent dye. Labeled samples were subsequently hybridized by loading onto a microarray slide using a Hybridization Chamber Kit part #G2534A (Agilent Technologies, Santa Clara, CA, USA) and Hybridization Gasket Slide Kit part #G2534-60003 (Agilent Technologies). Hybridization was performed over 16 h at 56 °C, followed by washing of the microarray slide as recommended by the manufacturer. Processed microarray slides were then scanned with an Agilent G2565CA Microarray Scanner System (Agilent Technologies, Santa Clara, CA, USA). Scanned images were imported using Agilent Feature Extraction software version 10.7.3.1 (Agilent Technologies, Santa Clara, CA, USA) and the fluorescence intensities of each image were quantified using the modified Exiqon (Vedbaek, Denmark) protocol and corresponding GAL files.
3
Comprehensive microRNA profiling using LNA array
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The miRCURY LNA TM microRNA Array (Exiqon, Denmark) was used. Three thousand and one hundred capture probes were employed, covering human, mouse, and rat microRNA in miRBase, as well as viral microRNA related to these species. Other reagents included the miRCURYTM Array Power Labeling kit (Cat #208032-A, Exiqon), 2X PCR master mix (Arraystar), RPMI1640 basic culture medium (Hyclone Co., cat: SH30022.03B), fetal bovine serum (FBS, GIBCO Co., cat: 16400-044), bispecific antibody (Prospec Co., cat: SV30010), Lipofectamine 2000(Invitrogen Co., cat: 11668-027), and real-time PCR kit (Thermo Co., cat: 11762-100), TRIZOL (Invitrogen Co., cat: 15596-026), and primer synthesis was done by Shanghai Sangon Biotech Co.
4
miRNA Expression Profiling Using LNA Array
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An miRCURY LNA TM MicroRNA Array (version 11.0; Exiqon) was used. Total RNA (1 μg) was labeled with an miRCURY LNA TM MicroRNA Power Labeling Kit (Exiqon) according to the instruction manual, and then the fluorescently labeled RNA was dissolved in hybridization buffer (350 μL) from the kit. The solution was incubated at 95 ° C for 2 min, slowly cooled down to room temperature, and applied to the array slide. The slide was further incubated at 56 ° C for 16 h, and washed with the washing buffer from the kit as described in the manual. Finally, the washed slide was dried, and scanned with a GenePix 4000B (Axon Instruments). The obtained data were normalized and analyzed using an Array-Pro Analyzer (version 4.5; Media Cybernetics) as reported previously [19] .
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