FFPE RNA/DNA Purification Kit
Catalog# : NB-25000
Price/Unit : $295.00
Unit : 50 preps
For the rapid isolation and purification of total RNA (including microRNA) and genomic DNA from FFPE tissue samples.
* Purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA)
* Preferential purification of RNA from other cellular components without the use of phenol or chloroform
* Purified RNA and genomic DNA are of the highest quality for use in numerous downstream applications
* Available in 96 well format
Bio-Synthesis's FFPE RNA/DNA Purification Kit provides a rapid method for the isolation and purification of total RNA (including microRNA) and genomic DNA from formalin-fixed paraffin-embedded (FFPE) tissue samples. Alternatively, the kit can be used to isolate total RNA alone or genomic DNA alone from FFPE tissue samples through varying the protease digestion time and performing optional RNase or DNAse digestions.
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. Bio-Synthesis's FFPE RNA/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.
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 and/or DNA will bind while most of the contaminants will be removed in the flowthrough. At this point, any traces of genomic DNA can be digested allowing for pure RNA samples to be isolated. Alternatively, traces of the RNA may be digested at this point if a pure sample of genomic DNA is required instead. The bound nucleic acid is then washed (WASH) in order to remove any impurities. Lastly, the purified total nucleic acd is eluted in 50 µL of the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart to the right.
This kit is also available in a 96-well format for high-throughput FFPE RNA/DNA purification. Purifictaion with the 96-well plate can be performed using either a vacuum manifold or centrifugation.
FEATURES AND BENEFITS
* High quality and integrity of the isolated nucleic acid - Both the purified total RNA and/or genomic DNA are of the highest quality and integrity, and can be used in any sensitive downstream application.
* Isolate a diversity of RNA species - All RNA species can be isolated, from large mRNA and ribosomal RNA down to microRNA and siRNA.
* High yields - Bio-Synthesis's FFPE RNA/DNA Purification Kit allows for the purification of high yields of total RNA or DNA.
* Rapid procedure - Isolate total RNA and/or genomic DNA from FFPE tissue sections using a rapid spin column format or a high-throughput 96-well plate format.
* Versatile procedure - The kit allows for the purification of either pure total RNA, pure genomic DNA or both RNA and genomic DNA through the use of tailored protease digestion times.
APPLICATIONS
The purified RNA and genomic DNA are of the highest available quality from FFPE samples, and can be used in a number of downstream applications including:
* qPCR
* qRT-PCR
* Reverse transcription PCR
* Primer extension
* Mutation screening
* Expression array assays
* Microarray analyses
* Sequencing
* Southern blotting
* SNP analysis
* Next Generation Sequencing
Figure 1. High Quality and Yield of Total RNA. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Bio-Synthesis's FFPE RNA/DNA 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.
FFPE RNA/DNA Purification Kit Contents
1. Digestion Buffer
2. Binding Solution
3. Enzyme Incubation Buffer
4. Wash Solution
5. Elution Buffer
6. Proteinase K
7. Spin Columns
8. Collection Tubes
9. Elution Tubes
10. Product Insert
Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. The Proteinase K should be stored in aliquots at -20°C 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.
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.
CleanAll RNA/DNA Clean-Up and Concentration Kit
Catalog# : NB-23800
Price/Unit : $240.00
Unit : 20 preps
For the rapid and efficient purification, cleanup and concentration of RNA or DNA
* Purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA)
* RNA is preferentially purified from other reaction components such as proteins and nucleotides, without the use of phenol or chloroform
* Purifies all sizes of DNA, from small PCR products to plasmids to genomic DNA.
* Rapid and efficient spin-column format
Features and Benefits | Specifications
BSI's CleanAll RNA/DNA Clean-Up and Concentration Micro Kit provides a rapid method for the purification, cleanup and concentration of RNA or DNA from different isolation methods or upstream applications. The kit can be used as an alternative to organic extraction and ethanol precipitation to clean up various enzymatic reactions. The CleanAll Kit purifies RNA from phenol/guanidine-based protocols or from various upstream enzymatic reactions such as DNase treatment, labeling and in vitro transcription. Furthermore, endotoxins can be removed from previously purified RNA solutions. The kit can be used to clean up DNA from digestions, ligations, PCR reactions, labeling reactions, DNA modification reactions and staining. The kit also provides a protocol for the rapid removal of endotoxins from previously purified DNA down to 0.1 EU/µg DNA or less.
Purification is based on spin column chromatography using BSI’s proprietary resin as the separation matrix. Briefly, the samples or reaction containing RNA or DNA is mixed with Binding Solution and the nucleic acids are bound to BSI's column (BIND). Under these conditions only the nucleic acids will bind to BSI's resin while most of the contaminating proteins and nucleotides are removed in the flowthrough. The bound nucleic acid is then washed to remove any remaining impurities (WASH). Lastly, the purified total RNA or DNA is eluted into 50 µL of the provided Elution Buffer or water (ELUTE). Please see procedure flowchart to the right.
FEATURES AND BENEFITS
* Versatile performance - Both DNA and RNA can be cleaned and concentrated using the same kit. Clean-up nucleic acids from a variety of isolation methods and upstream enzymatic reactions.
* Fast and easy processing - Rapid spin-column format allows for the processing of 10 samples in 20 minutes.
* Suitable for all sizes of RNA - Purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA).
* Suitable for all sizes of DNA - Purify all sizes of DNA, from small PCR products, native or linearized plasmids, to genomic DNA. Using an alternative protocol, even oligonucleotides and smaller DNA fragments can be purified.
* Optional endotoxin removal - Optional protocol is provided for the rapid removal of endotoxins from previously purified RNA or DNA.
* No phenol:chlorofom extractions - BSI’s CleanAll kit purifies RNA without the use of harmful chemicals such as phenol or chloroform
APPLICATIONS AND DATA
The kit can be used to clean up RNA and DNA from upstream applications such as:
* Digestions
* Ligations
* PCR reactions
* Labeling reactions
* DNA modification reactions
* Staining
Figure 1. High Recoveries and Efficient Endotoxin Removal. High recoveries of DNA can be seen when using the CleanAll kit, and these high recoveries are not compromised even during endotoxin removal. In Panel A, 1 µg of a PCR product was cleaned using the CleanAll RNA/DNA Clean-Up and Concentration Kit as well as BSI's PCR Purification Kit. More than 0.9 µg of the DNA was recovered from both kits with a comparable percent recovery of over 90%. In Panel B, 10 µg of plasmid DNA preparation was cleaned from endotoxin by using the CleanAll RNA/DNA Kit. Endotoxin levels of the plasmid DNA sample were reduced from 0.75 EU/µg plasmid DNA to less 0.05 EU/µg plasmid DNA. Approximately 95% of the endotoxin units in the DNA sample were removed using the kit, therefore the kit allows for high recoveries coupled with exceptional cleanup.
Figure 2: Clean-Up of RNA with High Recovery. BSI’s CleanAll RNA/DNA Clean-Up and Concentration Kit can be used to clean up various enzymatic reactions including DNase treatment. In Figure 2 above, Lane 1 is the RNA input, while lanes 2-5 contain the RNA that has been cleaned using BSI’s CleanAll Kit. It can be seen that in all cases the recovery is high, and the purified RNA is intact and of a high quality.
CleanAll RNA/DNA Clean-Up and Concentration Kit Components
1. Binding Solution
2. Wash Solution
3. Elution Buffer
4. Micro Spin Columns
5. Collection Tubes
6. Elution tubes
7. Product Insert
Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. 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.
FEATURES AND BENEFITS
Figure 1. High Quality and Yield of Total RNA. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Bio-Synthesis's FFPE RNA/DNA 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.
