FFPE RNA/DNA Purification Plus Kit Dx
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Sequential isolation and purification of total RNA and genomic DNA from FFPE tissue samples
Research version available here
Aperçu du produit
Norgen’s FFPE RNA/DNA Purification Plus Kit Dx provides a rapid method for the sequential isolation and purification of total RNA (including microRNA) and genomic DNA from formalin-fixed paraffin-embedded (FFPE) tissue samples for subsequent in vitro diagnostic use. Using formalin to fix tissues leads to crosslinking of the nucleic acids and proteins, and the process of embedding the tissue samples can also lead to fragmentation of the nucleic acids over time. Norgen’s FFPE RNA/DNA Purification Plus Kit Dx provides conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of nucleic acids. The kit is able to purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA), depending on the age of the FFPE tissue as the degree of fragmentation of the RNA will increase over time.
This kit is designed to be used with any downstream application employing enzymatic amplification or other enzymatic modifications of nucleic acids followed by signal detection or amplification. Any diagnostic results generated using the RNA and DNA isolated with Norgen’s FFPE RNA/DNA Purification Plus Kit Dx in conjunction with an in vitro diagnostic assay should be interpreted with regard to other clinical or laboratory findings.
To minimize irregularities in diagnostic results, suitable controls for downstream applications should be used.
Norgen’s FFPE RNA/DNA Purification Plus Kit Dx is intended for use by professional users such as technicians, physicians and biologists experienced and trained in molecular biological techniques.
Norgen’s FFPE RNA/DNA Purification Plus Kit Dx does not provide a diagnostic result. It is the sole responsibility of the user to use and validate the kit in conjunction with a downstream in vitro diagnostic assay.
Détails
Supporting Data
Figure 1. Superior Recovery of High Quality RNA and DNA from FFPE Spleen Tissues. Norgen's FFPE RNA/DNA Purification Plus Kit Dx isolates FFPE RNA and DNA that exceeds the yield of competitors. Total RNA and DNA was isolated from equal amount of hamster FFPE spleen sections (20 micron thickness) using Norgen's FFPE RNA /DNA Purification Plus Kit Dx and a leading competitor’s kits. Triplicate isolations were performed for each product. The top graphs demonstrate the mean yield of RNA (Panel A) and DNA (Panel B) according to NanoDrop measurement. The bottom graphs showed the mean 260:280 ratio and 260:230 ratio of RNA (Panel C) and DNA (Panel D) according to NanoDrop measurement. Norgen's kit consistently purified total RNA and DNA with a higher yield and higher quality than for those obtained using the market competitor's kits.
Figure 2. Recovery of High Quality DNA and True Total RNA including microRNA from Hamster Spleen. Genomic DNA isolated using Norgen's FFPE RNA/DNA Purification Plus Kit Dx is of high quality with effective qPCR amplification. Panel A showed that in a qPCR using primers against the 5S rRNA gene using equal amount of FFPE DNA as template, the DNA isolated with Norgen's kit showed better amplification (lower Ct value). Moreover, Norgen's FFPE RNA/DNA Purification Plus Kit isolates true total RNA including microRNA. With the same amount of RNA as template in RT-qPCR reactions for transcripts of miR-21 (Panel B for microRNA) and beta-Actin (Panel C for mRNA), RNA isolated by Norgen showed better amplification in beta-Actin and significantly better in miR-21 microRNA.
Kit Specifications
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Maximum Column Binding Capacity (RNA)
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35 μg
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Maximum Column Binding Capacity (gDNA)
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10 μg
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Maximum Column Loading Volume
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600 μL
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Size of RNA Purified
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All sizes, including small RNA (< 200 nt) |
Maximum Amount of Starting Material |
4 sections <20 μM thick
10 mg of unsectioned block |
Storage Conditions and Product Stability
The DNAse I and Proteinase K should be stored at -20°C upon arrival. All other solutions and kit components should be kept tightly sealed and stored at room temperature. All solutions and plastics can be used until the expiration date specified on their labels.
Component | Cat. 54300 (50 preps) |
---|---|
Digestion Buffer | 2 x 20 mL |
Binding Solution | 2 x 20 mL |
Enzyme Incubation Buffer A | 6 mL |
RNA Wash Solution | 24 mL |
DNA Wash Solution | 38 mL |
RNA Elution Solution | 6 mL |
DNA Elution Buffer | 15 mL |
Proteinase K | 2 x 12 mg |
DNase I | 200 μL |
RNA Purification Micro Columns | 50 |
DNA Purification Micro Columns | 50 |
Collection Tubes | 100 |
Elution Tubes (1.7 mL) | 100 |
Product Insert | 1 |