FFPE RNA/DNA Purification Plus Kit
Sequential isolation and purification of total RNA and genomic DNA from FFPE tissue samples
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For research use only and NOT intended for in vitro diagnostics.
FFPE RNA/DNA Purification Plus Kit
Sequential isolation and purification of total RNA and genomic DNA from FFPE tissue samples
Overview
- Fast and easy processing using rapid spin-column format
- High yields and quality of nucleic acids
- Separate fractionation of RNA and DNA
- Isolate total RNA, from large rRNA down to microRNA (miRNA)
- No phenol or chloroform extractions
- Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen's NGS services
- Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
Norgen’s FFPE RNA/DNA Purification Plus Kit 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. The RNA and DNA are sequentially purified into separate fractions using this kit. 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 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. The RNA is purified from other cellular components without the use of phenol or chloroform. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays. The purified genomic DNA is also of the highest quality, and can be used in PCR reactions, sequencing, Southern blotting and SNP analysis.
Details
Supporting Data
Figure 1. Superior Recovery of High Quality RNA and DNA from FFPE Spleen Tissues. Norgen's FFPE RNA/DNA Purification Plus Kit 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 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 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
|
|
Maximum Binding Capacity
|
Up to 35 μg RNA
Up to 10 μg DNA |
Maximum Column Loading Volume
|
600 μL
|
Size of RNA Purified
|
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
All solutions should be kept tightly sealed and stored at room temperature. The DNAse I and Proteinase K should be stored at -20°C upon arrival. This kit is stable for 1 year after the date of shipment.
Component | Cat. 54300 (50 preps) |
---|---|
Digestion Buffer A | 2 x 25 mL |
Buffer RL | 40 mL |
Enzyme Incubation Buffer A | 6 mL |
Wash Solution A | 2 x 38 mL |
Elution Solution A | 6 mL |
Elution Buffer F | 15 mL |
Proteinase K | 2 x 12 mg |
DNase I | 1 vial |
RNA Purification Micro Columns | 50 |
DNA Purification Micro Columns | 50 |
Collection Tubes | 100 |
Elution Tubes (1.7 mL) | 100 |
Product Insert | 1 |
Documentation
Citations
Title | Multiregion sequencing of sarcomatoid renal cell carcinoma arising from autosomal dominant polycystic kidney disease |
Journal | Molecular Genetics & Genomic Medicine. 2022. |
Authors | Elizabeth Lee, Peiyong Guan, Abner Herbert Lim, Jui Wan Loh, Grace Fangmin Tan, Tracy Loh, Dave Yong Xiang Ng, Jing Yi Lee, Shane Goh, Wei Liu, Cedric Chuan-Young Ng, Bin Tean Teh, Jason Yongsheng Chan |
Title | Decreased expression of miR-23b is associated with poor survival of endometrial cancer patients |
Journal | Research Square. 2022. |
Authors | Paweł K. Włodarski, Klaudia Klick, Tomasz M. Grzywa, Alicja Klinke, Aleksandra Mielniczuk, Jarosław Wejman, Joanna Ostrowska, Agata Gondek |
Title | Methodological Challenges of Digital PCR Detection of the Histone H3 K27M Somatic Variant in Cerebrospinal Fluid |
Journal | Pathol Oncology & Research. 2022. |
Authors | Zaytseva M, Usman N, Salnikova E, Sanakoeva A, Valiakhmetova A, Chervova A, Papusha L, Novichkova G, Druy A |
Title | The use of whole genome methylation scanning to define genes preferentially suppressed in African American Prostate Cancer |
Journal | Cancer Epidemiology, Biomarkers and Prevention. 2017. |
Authors | Farah Rahmatpanah, Kathleen McGuire, Michael Lilly, Michael McClelland and Dan Mercola |
Title | MicroRNA-34c-5p is related to recurrence in laryngeal squamous cell carcinoma |
Journal | The Laryngoscope. 2015. |
Authors | Massimo Re MD, Artan Çeka MD, Corrado Rubini MD, Luigi Ferrante PhD, Antonio Zizzi PhD, Federico M. Gioacchini MD, Michele Tulli MD, Liana Spazzafumo PhD, Stefano Sellari-Franceschini MD, Antonio D. Procopio MD, andFabiola Olivieri PhD |
Title | In-Vivo Fusion of Human Cancer and Hamster Stromal Cells Permanently Transduces and Transcribes Human DNA |
Journal | PLOS ONE. 2014. |
Authors | David M. Goldenberg, Robert J. Rooney, Meiyu Loo, Donglin Liu, Chien-Hsing Chang |
Title | The miRNA-23b/27b/24 Cluster Promotes Breast Cancer Lung Metastasis by Targeting Metastasis-Suppressive Gene Prosaposin. |
Journal | The Journal of Biological Chemistry. 2014. |
Authors | Brian Ell, Qiong Qiu, Yong Wei, Laura Mercatali, Toni Ibrahim, Dino Amadori and Yibin Kang |
Title | Tumor-Induced Osteoclast miRNA Changes as Regulators and Biomarkers of Osteolytic Bone Metastasis. |
Journal | Cancer Cell. 2013. |
Authors | Ell B, Mercatali L, Ibrahim T, Campbell N, Schwarzenbach H, Pantel K, Amadori D, Kang Y. |
Title | Tyrosine kinase receptor status in endometrial stromal sarcoma: an immunohistochemical and genetic-molecular analysis. |
Journal | International Journal of Gynecology Pathology. 2012. |
Authors | Cossu-Rocca P, Contini M, Uras MG, Muroni MR, Pili F, Carru C, Bosincu L, Massarelli G, Nogales FF, De Miglio MR. |
Title | Factors affecting the yield of microRNAs from laser microdissectates of formalin-fixed tissue sections. |
Journal | BMC Research Notes. 2012. |
Authors | Patnaik SK, Kannisto E, Yendamuri S. |
Title | Overexpression of the Lung Cancer-Prognostic miR-146b MicroRNAs Has a Minimal and Negative Effect on the Malignant Phenotype of A549 Lung Cancer Cells. |
Journal | PLoS One. 2011. |
Authors | Patnaik S, Kannisto E, Mallick R, Yendamuri S. |