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.
Rneasy serum kit
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.
Lab products found in correlation
2 protocols using rneasy serum kit
Profiling Serum microRNA via RNA-Seq
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.
Serum Analysis of Lymph Node Samples
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