FFPE DNA Purification Kit - for Research Use Add to Cart
• Fast and easy processing using rapid and convenient spin-columns
• Isolate high quality and high yield DNA
• DNA is free of inhibitors and ready for downstream use including SNP (single nucleotide polymorphism) and short-tandem repeat (STR) genotyping
This kit provides a rapid method for the isolation and purification of DNA from formalin-fixed paraffin-embedded (FFPE) tissue samples. 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 DNA Purification Kit provides conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of nucleic acids. The purified DNA is of high yield and integrity and is free of inhibitors, ready for use in a number of downstream applications including qPCR, mutation screening, microarray analysis, sequencing, single nucleotide polymorphism (SNP) and short-tandem repeat (STR) genotyping. The protocol can be completed in as little as 1 hour.
Figure 1. Comparison of Total DNA Yield Isolated by Norgen´s FFPE DNA Kit 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 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 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
Figure 4. Comparison of DNA molecular weight range isolated by Norgen´s FFPE DNA Kit 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.
- T.M. Zohoncon, T.C. Ouedraogo, L.V.C. Brun, D. Obiri-Yeboah, W.F. Djigma, S. Kabibou, S. Ouattara, M. Gomina, A.T. Yonli, V.J.T.E. Bazie, C. Ouedraogo, O. Lompo, S.A. Akpona and J. Simpore. (2016) Molecular Epidemiology of High-Risk Human Papillomavirus in High-Grade Cervical Intraepithelial Neoplasia and in Cervical Cancer in Parakou, Republic of Benin. Pakistan Journal of Biological Sciences