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RNA Isolation

Isolation from Blood (RNA)

Agencourt RNAdvance Blood

Built on Solid Phase Reversible Immobilization (SPRI) paramagnetic bead-based technology, the Agencourt RNAdvance Blood system is a ribonucleic acid (RNA) isolation process. It enables the life science community to purify high quality RNA from blood collected using PAXgene* tubes. The kit is formatted to process one 96-well plate in about 3.5 hours.

Overview

  • Produces high-quality nucleic acid
  • Compatible with downstream gene expression analysis techniques such as qRT-PCR and microarray
  • Amenable to automated processing on the Beckman Coulter Biomek FXP/NXP Span-8 Lab Automation Workstation
  • Vacuum filtration or organic extraction not required


Application: Total RNA extraction from blood
Downstream Application Techniques: Gene expression via qRT-PCR and microarray analysis
 


Process Overview
 


1. Lysis buffer added to the blood and mixed 2. Lysis and Proteinase K digestion 3. Addition of Bind 1 Buffer
4. Magnetic separation of beads from supernatant, wash with Wash Buffer and Ethanol 5. DNase I reaction
6. Rebinding with Bind 2 Buffer 7. Magnetic separation of beads from supernatant, wash with Wash Buffer and
Ethanol 8.Elution


Superior Yield and Quality 
 

In the study below (Figure 1), Agencourt RNAdvance Blood produced approximately 700 ng of RNA per 400 µL of PAXgene blood, which equates to almost 17 µg of RNA per tube of PAXgene-preserved blood. By using the SPRI paramagnetic bead-based process, Agencourt RNAdvance Blood recovered over 30% more RNA than the PAXgene Blood RNA kit, while demonstrating consistent purity.
 

Figure 1.  Average RNA yield (in µg/mL) from 24 400 µL PAXgene-preserved blood samples extracted with the Agencourt RNAdvance Blood kit and 9.4 mL of PAXgene-preserved blood extracted with the PAXgene Blood RNA kit. Average 260/280 ratios across the 24 extraction performed using Agencourt RNAdvance Blood was 1.8 with a Standard Deviation of 0.13 and %CV of 7.24. The elution volume for the Agencourt RNAdvance Blood extraction was 20 µL.

Webinars & Symposiums

  • High-Performance RNA & miRNA Extractions using SPRI

    Bee Na Lee, PhD, Applications Scientist Beckman Coulter, Inc.

    Click to play  

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