FFPE RNA Purification Kits
For the rapid and efficient extraction and purification of RNA (including microRNA) from FFPE samples
![FFPE RNA Purification Kit](/sites/default/files/styles/norproduct_imagekit/public/images/product_images/25300-FFPE-RNA-Purification-Kits-WEB.jpg?itok=UkrLmYFH)
For research use only and NOT intended for in vitro diagnostics.
CEIVDD marked diagnostic version available here
FFPE RNA Purification Kits
For the rapid and efficient extraction and purification of RNA (including microRNA) from FFPE samples
Overview
- Extract total RNA (including microRNA) from FFPE samples
- No phenol extraction step
- Includes DNase for optional on-column DNA removal
- Isolated RNA is of the highest quality and integrity
- Isolate a diversity of RNA species
- 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 Purification Kits provide a rapid method for the isolation and purification of total RNA (including microRNA) from formalin-fixed paraffin-embedded (FFPE) tissue samples in as little as 1 hour. Using formalin to fix tissues leads to crosslinking of the RNA and proteins, and the process of embedding the tissue samples can also lead to fragmentation of the RNA over time. Norgen’s FFPE RNA Purification Kits provide conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of RNA. These kits are 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 fragmentation of the RNA is known to occur over time. The RNA is preferentially purified from other cellular components without the use of phenol or chloroform.
FFPE RNA Purification Kit (Spin Column)
Maximum loading volume of 650 μL per column, and a maximum binding capacity of 50 μg per column.
FFPE RNA Purification 96-Well Kit (High Throughput)
Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. Purification can be performed using either a vacuum manifold or centrifugation. Maximum loading volume of 400 μL per well, and a maximum binding capacity of 50 μg per well.
Details
Supporting Data
Figure 1. High Quality and Yield of Total RNA. Norgen's FFPE RNA Purification Kit isolates FFPE RNA that exceeds both yield and quality of competitors. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Norgen's FFPE RNA Purification Kit and a leading competitor’s kit. One microliter of the 50 µL purified RNA was then resolved on an Agilent 2100 BioAnalzyer using an RNA Nano 6000 chip. As it can be seen, Norgen not only isolated higher yields of total RNA, but the RNA was also of a higher quality as evidenced by the higher RIN values obtained with Norgen's RNA.
Figure 2. Higher Yield of FFPE RNA Isolated by Norgen's FFPE RNA Purification Kit. Norgen's FFPE RNA Purification Kit isolates FFPE RNA that exceeds the yield of competitors. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Norgen's FFPE RNA Purification Kit and two leading competitor's kits. Eighteen isolations were performed for each product. The graph demonstrates the mean yield of RNA according to spectrophotometry for 18 sample replicates. Vertical bars represent the standard deviation. Norgen's kit consistently purified total RNA with a higher yield than for those obtained using the market competitor's kits.
Figure 3. High Yield of a Diversity of RNA Species. Norgen's FFPE RNA Purification Kit effectively recovers all sizes of RNA from large mRNA to small RNA including microRNAs. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Norgen's FFPE RNA Purification Kit and a leading competitor’s kit. Five microliters of the 50 µL purified RNA was then used as the template in a 20 µL reverse transcription reaction with oligo dT primer or miR-21 stem-loop reverse primer. Two microliters of the RT reaction was then used in a 15 µL qPCR reaction for detecting the beta-actin gene (Panel A) and for detecting miR-21(Panel B), respectively. In both graphs the blue lines correspond to Norgen isolated-RNA and the green lines correspond to competitor-isolated RNA. As it can be seen, Norgen's kit isolated higher yields of RNA in both cases, as indicated by the lower Ct values of the blue lines. Also, Norgen's kit successfully isolated not only large RNA (Panel A) but also microRNA (Panel B), indicating the diversity of RNA species isolated.
Figure 4. Excellent Performance in Downstream Application such as RT-qPCR. Norgen's FFPE RNA Purification Kit effectively recovers all sizes of RNA, from large mRNA to small RNA including microRNA, that perform effectively in sensitive applications such as RT-qPCR. Total RNA was isolated from equal amounts of an FFPE tissue sample using Norgen's FFPE RNA Purification Kit and two market competitors. The graph demonstrates the mean delta Ct value for 18 sample replicates. Vertical bars represent the standard deviation. Norgen's kit consistently isolates RNA of a higher yield and quality than that obtained by the two leading market competitors as indicated by the larger delta Ct values. Not only were the large RNA species detected including rRNA and mRNA, but small RNA species (microRNA) were also detected from the sample, indicating the diversity of RNA species isolated.
Figure 5. High Yield and Consistent Recovery of a Diversity of RNA Species. Norgen's FFPE RNA Purification 96-Well Kit effectively recovers all sizes of RNA including large mRNA to small RNA including microRNAs. Total RNA was isolated from two equal amounts of an FFPE tissue sample using Norgen’s FFPE RNA Purification Kit. The purified RNA was then used as the template in a RT-qPCR for detecting the beta-actin gene (Panel A) and for detecting miR-21 (Panel B). As it can be seen, Norgen's kit successfully and consistently isolated not only large RNA (Panel A) but also microRNA (Panel B), indicating the diversity of RNA species isolated.
Kit Specifications
|
|
Maximum Column Binding Capacity |
Up to 50 µg RNA
|
Maximum Loading Volume Per Spin Column |
650 µL
|
Size of RNA Purified |
All sizes, including small RNA (< 200 nt)
|
Time to Complete 10 Purifications |
1-4 hours*
|
Maximum Amount of Starting Material |
5 slices of < 20 µm thick paraffin slices
25 mg of unsectioned block |
Average Yield |
Variable due to age of paraffin blocks
~2-3 µg of Total RNA per 1 mg of fresh FFPE hamster kidney |
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 from the date of shipment.
Component | Cat. 25300 (50 preps) | Cat. 25400 (2 x 96 preps) |
---|---|---|
Digestion Buffer A | 25 mL | 2 x 25 mL |
Buffer RL | 30 mL | 2 x 30 mL |
Enzyme Incubation Buffer | 6 mL | 2 x 6 mL |
Wash Solution A | 38 mL | 2 x 38 mL |
Elution Solution A | 6 mL | 2 x 20 mL |
Proteinase K | 12 mg | 2 x 20 mg |
DNase I | 1 vial | 2 x 500μL |
Micro Spin Columns | 50 | - |
96-Well Incubation Plate | - | 2 |
96-Well Plate | - | 2 |
Adhesive Tape | - | 8 |
Collection Tubes | 50 | - |
96-Well Collection Plate | - | 2 |
Elution Tubes (1.7 mL) | 50 | - |
96-Well Elution Plate | - | 2 |
Product Insert | 1 | 1 |
Documentation
Citations
Spin Column (25300)
Title | Ketogenic diet alters the epigenetic and immune landscape of prostate cancer to overcome resistance to immune checkpoint blockade therapy |
Journal | Cancer Research. 2024. |
Authors | Sean Murphy, Sharif Rahmy, Dailin Gan3, Guoqiang Liu1, Yini Zhu1, Maxim Manyak, Loan Duong, Jianping He, James H Schofield, Zachary T Schafer, Jun Li, Xuemin Lu, Xin Lu |
Title | Identification of Differentially Expressed Intronic Transcripts in Osteosarcoma |
Journal | Non-Coding RNA. 2022. |
Authors | Emel Rothzerg, Jiake Xu, and David Wood |
Title | Zona Pellucida sperm-binding protein 3 receptor distribution during Gopc−/− globozoospermic spermatogenesis |
Journal | Microscopy Research & Technique. 2021. |
Authors | Maider Bizkarguenaga,Laura Gomez-Santos,Juan Francisco Madrid,Francisco José Sáez,Edurne Alonso |
Title | The impact of RNA extraction method on accurate RNA sequencing from formalin-fixed paraffin-embedded tissues |
Journal | BMC Cancer. 2019. |
Authors | Michal Marczyk, Chunxiao Fu, Rosanna Lau, Lili Du, Alexander J. Trevarton, Bruno V. Sinn, Rebekah E. Gould, Lajos Pusztai, Christos Hatzis & W. Fraser Symmans |
Title | miR-17-92a-1 cluster host gene (MIR17HG) evaluation and response to neoadjuvant chemoradiotherapy in rectal cancer |
Journal | OncoTargets and Therapy. 2016. |
Authors | Molinari C, Salvi S, Foca F, Teodorani N, Saragoni L, Puccetti M, Passardi A, Tamberi S, Avanzolini A, Lucci E, Calistri D |
Title | Involucrin Expression and Association with Ki-67 in Paraffin Embedded Tissue of Canine Skin Tumors: A Retrospective Study |
Journal | The Thai Journal of Veterinary Medicine. 2016. |
Authors | Assawawongkasem, N., Suriyaphol, G., Srisuwatanasagul, S., Theerawattanasirikul, S., & Sailasuta, A |
Title | Correlative Analysis of miRNA Expression and Oncotype Dx Recurrence Score in Estrogen Receptor Positive Breast Carcinomas |
Journal | PLOS ONE. 2015. |
Authors | Rajyasree Emmadi , Emanuele Canestrari, Zarema H. Arbieva, Wenbo Mu, Yang Dai, Jonna Frasor, Elizabeth Wiley |
Title | Ocoxin® oral solution slows down tumor growth in an experimental model of colorectal cancer metastasis to the liver in Balb/c mice. |
Journal | Oncology Reports. 2015. |
Authors | Márquez J, Mena J, Hernandez-Unzueta I, Benedicto A, Sanz E, Arteta B, Olaso E |
Title | MicroRNA molecular profiling from matched tumor and bio-fluids in bladder cancer |
Journal | Molecular Cancer. 2015. |
Authors | David A. Armstrong, Benjamin B. Green, John D. Seigne, Alan R. Schned and Carmen J. Marsit |
Title | Telomeric repeat-containing RNA (TERRA) constitutes a nucleoprotein component of extracellular inflammatory exosomes |
Journal | PNAS. 2015. |
Authors | Zhuo Wang, Zhong Deng, Nadia Dahmane, Kevin Tsai, Pu Wang, Dewight R. Williams, Andrew V. Kossenkov, Louise C. Showe, Rugang Zhang, Qihong Huang, José R. Conejo-Garcia, and Paul M. Lieberman |
Title | A phase II and pharmacodynamic study of sunitinib in relapsed/refractory oesophageal and gastro-oesophageal cancers |
Journal | British Journal of Cancer. 2015. |
Authors | C Wu, S Mikhail, L Wei, C Timmers, S Tahiri, A Neal, J Walker, S El-Dika, M Blazer, J Rock, D J Clark, X Yang, J L Chen, J Liu, M V Knopp and T Bekaii-Saab |
Title | MiR-205 and MiR-375 MicroRNA Assays to Distinguish Squamous Cell Carcinoma from Adenocarcinoma in Lung Cancer Biopsies |
Journal | Journal of Thoracic Oncology. 2015. |
Authors | Patnaik, Santosh, Mallick, Reema, Kannisto, Eric MS, Sharma, Rohit, Bshara, Wiam, Yendamuri, Sai M, Dhillon, Samjot Singh |
Title | LMP1-deficient Epstein-Barr virus mutant requires T cells for lymphomagenesis |
Journal | The Journal of Clinical Investigation. 2014. |
Authors | Shi-Dong Ma, Xuequn Xu, Julie Plowshay, Erik A. Ranheim, William J. Burlingham, Jeffrey L. Jensen, Fotis Asimakopoulos, Weihua Tang, Margaret L. Gulley, Ethel Cesarman, Jenny E. Gumperz and Shannon C. Kenney |
Title | Robust global microRNA expression profiling using next-generation sequencing technologies. |
Journal | Laboratory Investigation. 2014. |
Authors | Tam S, Borja R, Tsao M, and McPherson J. |
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. |
High Throughput (25400)
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. |