RNAdvance Tissue Performance and Data
Total RNA Extraction from Tissues
The RNAdvance Tissue kit is a ribonucleic acid (RNA) isolation reagent kit built on SPRI paramagnetic bead-based technology. It enables extraction of total RNA from a wide variety of tissues without the hazards and waste removal issues of organic solvents or the time and labor consuming steps of vacuum filtration and centrifugation procedures. The extraction can be run manually in a 2 mL tube format or 96-well format or automated in 96-well format on variety of Beckman Coulter Biomek liquid handling workstations. This extraction process produced higher recovery of total RNA than traditional column-based approaches in the experiments conducted for data shown below.
- Compatible with downstream PCR based applications
- Extraction and purification of high quality RNA from many tissue types
- Efficient removal of genomic DNA and other contaminants
High Recovery of RNA from a Variety of Tissue Types Without Sacrificing Purity
RNA was extracted from 10mg of brain, liver, lung, kidney or tail tissues using the RNAdvance Tissue and other commercially available kits. In Figure 1 (above) RNA yield was quantified using the NanoDrop (Thermo Fisher Scientific). RNAdvance Tissue recovered higher amounts of RNA from the five different tissue types used than the other commercially available kits. In Figure 2 (above) samples were assessed for purity using the NanoDrop (Thermo Fisher Scientific). For all tissue types, RNAdvance tissue purified RNA with satisfactory A260/280 ratios.
Remarkable RNA Quality
RNA extracted from 10mg of brain tissue using RNAdvance Tissue was run on the Agilent 2100 Bioanalyzer Eukaryote Total RNA Nano chip. In Figures 3 and 4 below, two traces of RNA extracted from brain tissue are shown. The RIN scores were >9 indicating that high quality and intact RNAs were recovered.
RNAdvance Tissue Provides RNA Suitable for Downstream PCR Applications
PCR amplifiability was assessed via qRT-PCR using a primer set (forward primer 5´- gacttcgagcaagagatgg-3´ and reverse primer 5´-agcactgtgttggcgtacag-3´) designed to span Exon 4 and 5 of the beta (β)-actin gene (ActB) to produce 327 base pair amplicons. In Figure 5 (below) the no RT control also demonstrates the removal of DNA that can interfere with downstream RNA applications. In Figure 6 (below) the RNA isolated using the RNAdvance Tissue kit was amplifiable indicating that the kit removed PCR inhibitors. These results indicate the usability of the RNA for downstream PCR applications.
Extract RNA from Samples in Less Time and with Less Pipette Actions Compared to users of Column-Based Kits
Figure 7 (below) represents total time to extract RNA for 1 to 96 samples using RNAdvance Tissue kit or a column based kits. At 20 samples total time to extract RNA from tissues is faster using RNAdvance Tissue kit. In Figure 8 (below) the total number of pipette actions, which include dispensing in a sample, mixing a sample, and discarding tips, required for 1, 8, 24, 48, and 96 samples. With the ability to use a multichannel pipette there is significantly less pipette actions at greater than 8 samples that need to take place than with column based kits.
RNAdvance Tissue Visual Workflow
- Lyse tissue in Lysis Buffer and Proteinase K
- Bind RNA to magnetic beads
- Separate magnetic beads from contaminants
- Wash magnetic beads with Wash Buffer and 70 % ethanol to remove contaminants
- Treat samples with DNase I
- Rebind RNA to magnetic beads with Wash Buffer
- Wash magnetic beads with 70% ethanol to remove contaminants
- Elute RNA from magnetic beads
- Transfer to new plate for storage
For Use in Manual or Automated Methods Based on Batch Size or Overall Throughput
Estimated hands-on-time and total time in hours required to perform 8, 24 and 96 RNAdvance Tissue RNA extractions. The extraction can be performed either manually or automated on a liquid handling system. The automated times reflect extraction performed on a Biomek i5 Multichannel 96 Genomics Workstation. Difference in time between manual and automation is indicated (NR=not recommended).
| RNAdvance Tissue | Manual | Automated | ||
|---|---|---|---|---|
| Batch Size | 8 | Hands-on-Time | 0.08 | 0.25 |
| Total Time | 1.00 | 2.50 | ||
| 24 | Hands-on-Time | 0.11 | 0.25 | |
| Total Time | 1.10 | 2.50 | ||
| 96 No DNase Treatment | Hands-on-Time | NR | 0.25 | |
| Total Time | NR | 2.00 | ||
| 96 | Hands-on-Time | NR | 0.25 | |
| Total Time | NR | 2.50 | ||
RNAdvance Tissue Products for RNA Extraction from Tissue
RNAdvance Tissue is available in three kit sizes based on your throughput needs. Click on the specific part number below for purchasing or request a quote. For more information check out our RNA isolation from tissue page featuring RNAdvance Tissue.
| PART # | NAME | PREPS |
|---|---|---|
| A32645 | RNAdvance Tissue Kit | 50 |
| A32649 | RNAdvance Tissue Kit | 96 |
| A32646 | RNAdvance Tissue Kit | 384 |
Not intended or validated for use in the diagnosis of disease or other conditions.
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