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Rneasy serum kit

Manufactured by Qiagen
Sourced in United Kingdom, United States

The RNeasy Serum kit is a laboratory equipment designed for the purification of total RNA from serum or plasma samples. It utilizes a silica-membrane technology to efficiently capture and purify RNA molecules from the sample, enabling their subsequent analysis and applications.

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2 protocols using rneasy serum kit

1

Profiling Serum microRNA via RNA-Seq

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Approximately the same amount of joint tissue per donor was pulverised into a powder with a dismembranator (Mikro-S, Sartorius, Melsungen, Germany) under liquid nitrogen, and total RNA was extracted using a miRNeasy kit (Qiagen, Crawley, UK).
Total RNA was extracted from 500 ​μl serum using a RNeasy Serum kit (Qiagen, Crawley, UK) with DNase treatment (Qiagen, Crawley, UK). RNA integrity (RIN) was confirmed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, USA). Ribosomal RNA (rRNA) was depleted using the Ribo-Zero™ rRNA Removal Kit (Epicentre, Madison, USA). Quality control prior to sequencing was performed by Qubit and Bioanalyzer (Agilient Technologies, Santa Clara, USA) analysis with RNA Pico chips and small RNA chips to measure RNA quantity, RNA integrity and calculate microRNA percentage. 100 ​ng of rRNA-depleted RNA per sample were submitted for library preparation using a NEB small RNA library kit (New England Biolabs (NEB), Ipswich, USA) with the addition of tobacco acid pyrophosphatase (Epicentre, Madison, USA). Pooled samples were size selected (120-300bp) and purified with Ampure beads (Agencourt, Beckman-Coulter, High-Wycombe, UK). Sequencing was undertaken on an Illumina HiSeq 2000 platform (Illumina, San Diego, USA) using 100 base paired-end reads.
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2

Serum Analysis of Lymph Node Samples

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Peripheral blood samples were centrifuged at 1,500 ×g for 10 min at 4 ℃. Serum was separated into EP tubes and stored at −80 ℃ until further use. The serum samples were divided into three comparison groups: 4 LNs vs. 4 HIs, 4 LNs vs. 6 NLNs, and 31 LNs vs. 62 NLNs vs. 72 HIs. The first two groups were used for next-generation sequencing analysis. A total of 165 samples (31 LNs vs. 62 NLNs vs. 72 HIs) were used to verify the selected candidate biomarker (Figure 1). Total RNA was extracted from serum using a TRIzol Kit (Invitrogen, Life Technologies, Carlsbad, CA, USA) and an RNeasy Serum Kit (Qiagen, Hilden, Germany) following the manufacturers’ instructions. The total RNA quantity and purity were confirmed with an Agilent High Sensitivity DNA Kit (Agilent Technologies, Inc., USA) using a Bioanalyzer 2100 (Agilent Technologies, Inc., USA).
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