EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Kit
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Detail
Overview
Ensure optimal results for sensitive applications like NGS
Up to a tenfold increase in microRNA mapping during sequencing runs to reduce costs
The purified exosomal RNA is free from any circulating RNA-binding proteins.
No phenol extractions, Proteinase K treatment, nor carrier RNA.
No time-consuming ultracentrifugation, filtration nor special syringes are required.
Concentrate isolated RNA into a flexible elution volume ranging from 50 μL to 100 μL.
Purification is based on EXTRAClean spin column chromatography that uses Norgen’s proprietary separation matrix.
Norgen’s EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Kit constitutes an all-in-one system for the purification of exosomes and the sequential isolation of RNA and free-circulating RNA from different plasma/serum sample volumes ranging from 50 μL and up to 10 mL. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS. The kit is designed to isolate all sizes of RNA, including microRNA. The kit provides a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or any protease treatments. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 μL to 100 μL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA can be used in a number of downstream applications including real time PCR, NGS application, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume
50-100 μL
Time to Complete 10 Purifications
40-45 minutes
Average Yields
Variable depending on specimen
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
Other Products
Cryptosporidium TaqMan RT-PCR Detection Kits
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Product Info
Overview
Detection kits for Cryptosporidium
CE-IVD marked version available for in vitro diagnostic use
Available in TaqMan format for analysis
Cryptosporidium is a parasite found in water that causes an infection in mammals termed cryptosporidiosis. It is one of the most common water-borne diseases and is found world-wide. It affects the intestines of mammals and typically causes an acute short-term infection. The most common symptom is self-limiting diarrhea in healthy individuals, however in immunocompromised individuals the symptoms are particularly severe and often fatal. There is no specific treatment for cryptosporidiosis other than fluid rehydration and management of any pain. Therefore early detection of Cryptosporidium in water is the foremost action to prevent the infection.
Cryptosporidium TaqMan RT-PCR Kit, 100 reactions
Ready to use format, including Master Mix for the target and PCR control to monitor for PCR inhibition and validate the quality
Specific Primer and Probe mix for the pathogen/virus/viroid of interest
Primer and Probe mix
Positive and negative control to confirm the integrity of the kit reagents
Cryptosporidium TaqMan RT-PCR Probe/Primer Set and Controls, 100 reactions
Specific Primer/Probe mix and Positive Control for the pathogen/virus/viroid of interest
Nuclease-free water
Can be used together with Norgen’s RT-PCR Master Mix (#28113) or customer supplied master mix
Storage Conditions and Product Stability All kit components can be stored for 1 year after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
Bioprocessing with Salt Active Nucleases – High Salt Conditions
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Product Info
Bioprocessing with Salt Active Nucleases – High Salt Conditions
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For SAN HQ, SAN HQ ELISA Kit, and now SAN HQ GMP
SAN HQ GMP is biochemically identical to SAN HQ but produced under GMP conditions.
Applications
Purification of biologics from residual nucleic acids in biopharma manufacturing
Purification of recombinant proteins and enzymes for research and diagnostic use
Removal of unwanted nucleic acids contamination in molecular biology reagents in challenging conditions
Reduction of viscosity in biological samples during production and automation
Vaccine manufacturing and viral vector preparation
DNA removal in high-salt lysates
SAN HQ – Peak performance at high salt conditions
Salt Active Nuclease High Quality (SAN HQ) is a Bioprocessing Grade nuclease developed as the most efficient solution for removal of both single and double stranded DNA and RNA at high salt conditions.
This nonspecific endonuclease has peak activity at salt concentrations between 400 – 700 mM (Fig. 1)
Non-enveloped viruses like Adenoviruses and Adeno-Associated Viruses (AAV’s) are inherently more robust with two distinct advantages: 1) They exhibit higher tolerance to additives like salt and detergents and 2) their production often involves the lysis of host cells, allowing for harvesting non-secreted vectors.
For Adeno-Associated Viruses (AAVs), which are often harvested from crude cell lysate, the high salt tolerance of SAN HQ is particularly beneficial. Salt is typically added to such lysates to reduce viral aggregation, facilitating more effective nuclease action to digest residual DNA.
SAN HQ’s is engineered for optimum activity in these high salt environments ensuring that you achieve unparalleled DNA removal without compromising the integrity of these robust viral vectors.
Key Benefits
Optimized Residual DNA Removal: Ensures efficient degradation of residual DNA in high-salt conditions, meeting stringent quality requirements for biologics and vaccines.
Boosted AAV Vector Purification: Enhances the purification process for adeno-associated viral vectors in high-salt conditions, improving quality and yield.
Streamlined Workflow: Eliminates the need for desalting stages, simplifying the bioprocessing protocol and saving time and resources.
Enable High-Throughput Processes: Facilitates scale-up and automation by working effectively in high-salt environments, increasing operational throughput.
Potential Surge in Virus Yield: Operates under conditions that may boost the titer yield of AAV production, potentially enhancing overall viral yield.
Economized Enzyme Usage: Reduces the need for excess enzyme and additional process adjustments, resulting in significant cost savings.
Minimized Risk of Process Disruptions: Offers reliable performance in various high-salt bioprocessing conditions, reducing the likelihood of disruptions due to enzyme inhibition.
Reliability: Provides consistent enzyme activity in challenging high-salt conditions, adding a layer of predictability and dependability to your operations.
Broader Applicability: Versatile enough to be used in a wide range of viral vector systems, expanding your research and production capabilities.
Enhanced Viral Stability: High-salt levels stabilize viral vectors, and SAN HQ operates effectively in these conditions, maintaining high yield and quality.
Host Cell Lysis: Facilitates efficient lysis of host cells in high-salt conditions, optimizing the harvest of both secreted and non-secreted viral vectors.
Key Features
High purity (≥ 98%)
No protease detected
Supplied with extended product documentation
Compatible with SAN HQ ELISA
The Challenge in Removing Host Cell Chromatin Impurities
In bioprocessing, the primary role of a nuclease is to efficiently digest and fragment host-cell DNA into sufficiently small pieces, facilitating its removal during downstream processing. While most nucleases can effectively degrade naked DNA into tiny fragments under optimal conditions—as demonstrated by M-SAN HQ and SAN HQ, which can digest dsDNA into fragments smaller than 6 nt—the reality in bioprocessing is more complex. (See fig. 5)
The DNA targeted for removal often exists as chromatin, embedded in a complex matrix containing remnants of the lysed host cell as well as large amounts of the therapeutic product.The product may or may not have an affinity for the chromatin you aim to remove.
High salt is often applied to mitigate issues like aggregation. The real challenge lies in a nuclease’s ability to efficiently fragment chromatin under these more complicated, high-salt, conditions—not merely degrading naked DNA under ideal circumstances.
SAN HQ ELISA kit is developed for the detection and quantification of SAN HQ and SAN HQ GMP. The kit is designed as a classical sandwich ELISA, with two monoclonal antibodies specific towards SAN HQ nuclease (fig 6).
Features
Sensitive: 0.4 – 25.6 ng/ml
Precise: RSD ≤ 15%
Accurate: 100% ± 15%
Stability: 12 months when stored between +2°C to +8°C
Document
For SAN HQ, SAN HQ ELISA Kit, and now SAN HQ GMP
SAN HQ GMP is biochemically identical to SAN HQ but produced under GMP conditions.
