FFPE RNA Purification Kit

FFPE RNA Purification Kit

Catalog# : NB-25300
Price/Unit : $320.00
Unit : 50 preps

For the rapid and efficient extraction and purification of RNA (including microRNA) from FFPE samples

* 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

Bio-Synthesis's FFPE RNA Purification Kit provides 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. Bio-Synthesis's FFPE RNA Purification Kit provides conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of RNA. 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 fragmentation of the RNA is known to occur over time.

Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. The process first involves deparaffinization of the FFPE samples through a series of xylene and ethanol washes. Next, the FFPE samples are digested with the provided Proteinase K and Digestion Buffer. Binding Solution and ethanol are then added to the lysate, and the solution is loaded onto a spin-column (BIND). Under these conditions only the RNA will bind while most of the contaminants will be removed in the flowthrough. At this point, any remaining traces of genomic DNA can be digested using an optional protocol, allowing for pure RNA samples to be isolated. The bound RNA is then washed (WASH) in order to remove any impurities. Lastly, the purified total RNA is eluted in 50 µL of the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart to the right.

FEATURES AND BENEFITS

* High quality and integrity of the isolated RNA - The purified total RNA is of the highest quality and integrity, and can be used in any sensitive downstream applications.
* Isolate a diversity of RNA species - All RNA species can be isolated, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA).
* High yields - Bio-Synthesis's FFPE RNA Purification Kit allows for the purification of high yields of total RNA.
* No phenol:chloroform extractions - Total RNA is isolated from FFPE tissue samples without the use of harmful chemicals such as phenol or chloroform.
* Rapid procedure - Isolate total RNA from FFPE tissue sections using a rapid spin column format in as little as 1 hour

APPLICATIONS

Purified RNA is of the highest quality, and can be used in a number of downstream applications including:

* Bioanalyzer
* Quantitative, real-time RT-PCR for both large mRNA and small RNA including miRNA
* RT-PCR for both large mRNA and small RNA including miRNA
* Northern blotting
* RNase protection
* Primer extension
* Expression array assays
* Next Generation Sequencing
* microRNA Cloning

Figure 1. High Quality and Yield of Total RNA. Bio-Synthesis'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 Bio-Synthesis'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, Bio-Synthesis's 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 Bio-Synthesis's RNA.

Figure 2. Higher Yield of FFPE RNA Isolated by Bio-Synthesis's FFPE RNA Purification Kit. Bio-Synthesis'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 Bio-Synthesis'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. Bio-Synthesis'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. Bio-Synthesis'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 Bio-Synthesis'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 Bio-Synthesis's isolated-RNA and the green lines correspond to competitor-isolated RNA. As it can be seen, Bio-Synthesis's kit isolated higher yields of RNA in both cases, as indicated by the lower Ct values of the blue lines. Also, Bio-Synthesis's kit successfully isolated not only large RNA (Panel A) but also microRNA (Panel B), indicating the diversity of RNA species isolated.

FFPE RNA Purification Kit Contents - Spin Columns
1. Digestion Buffer
2. Binding Solution
3. Enzyme Incubation Buffer
4. Wash Solution
5. RNA Elution Buffer
6. Proteinase K
7. DNase I
8. Spin Columns
9. Collection Tubes
10. Elution Tubes
11. Product Insert
Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. The DNAse I should be stored at -20°C upon arrival. The Proteinase K should be stored in aliquots at -20oC upon reconstitution. These reagents should remain stable for at least 1 year in their unopened containers.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.