Features and Benefits

Norgen’s FFPE DNA Purification Kit Dx provides a rapid method for the isolation and purification of 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 DNA over time. Norgen’s FFPE DNA Purification Kit Dx provides conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of DNA. Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. The DNA is preferentially purified from other cellular components without the use of phenol or chloroform.

This kit is designed to be used with any downstream application employing enzymatic amplification or other enzymatic modifications of DNA followed by signal detection or amplification. Any diagnostic results generated using the DNA isolated with Norgen’s FFPE DNA Purification 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 DNA Purification 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 DNA Purification 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.

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Supporting Data

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Figure 1. Comparison of Total DNA Yield Isolated by Norgen's FFPE DNA Kit Dx and a Leading Competitor FFPE DNA Purification Kit. DNA was isolated from 10 mg of FFPE kidney blocks. DNA concentrations were measured using the NanoVue spectrophotometer (GE Healthcare). Norgen's kit was found to have a much higher DNA yield, compared to the competitor kit.

Figure 2. Comparison of DNA Quality Isolated by Norgen's FFPE DNA Kit Dx and a Leading Competitor FFPE DNA Purification Kit. DNA was isolated from 10 mg of FFPE kidney blocks. Quality was assessed using A260:A280 and A260:A230 ratios generated from the NanoVue spectrophotometer (GE Healthcare). While Norgen and the competitor kit were found to have similar A260:A280 ratios, Norgen was found to have a much higher A260:A230 ratio, indicating higher quality DNA.

Figure 3. Comparison of qPCR Performance from FFPE DNA Isolated by Norgen's FFPE DNA Kit Dx and a Leading Competitor FFPE DNA Purification Kit. Total DNA was isolated from 10 mg of FFPE kidney blocks, in triplicate. A qPCR reaction was performed using 500 ng of purified DNA in a 2 µL reaction, using Norgen's 2x qPCR Mastermix (Cat# 28007) spiked with SYBR Green (Bio-Rad). The reaction took place in the iCycler iQ™Real-Time PCR Detection System (Bio-Rad), using beta-actin primers. The graph depicts the average Ct value generated from DNA isolated from both kits. Norgen and the leading competitor were found to isolate DNA from FFPE tissues with the highest quality, performing similarly in a qPCR reaction.

Figure 4. Comparison of DNA molecular weight range isolated by Norgen's FFPE DNA Kit Dx and a Leading Competitor FFPE DNA Purification Kits. DNA was isolated from 10 mg of FFPE kidney blocks. Based on the intensity of the bands, Norgen's kit captured more total DNA. Moreover, the DNA isolated by Norgen's kit covered a larger MW size compared with that isolated by the competitor kit. Both very high molecular DNA (intact genomic DNA, black arrow) as well as small molecular weight DNA were recovered. M = Norgen's HighRanger Molecular Weight Ladder.

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