HCM011 Selenite Cystine Broth (SC)

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.

See Case Study on Efficient Fragmentation: Superior Chromatin Digestion Before Flow-Through Purification

SAN HQ ELISA Kit

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).

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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

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.

Overview

• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias
• Versatile sample input ranges
• Isolate all sizes of free-circulating RNA, including microRNA​
• Bind and elute all RNA irrespective of size or GC content, without bias
• The purified exosomal RNA is free from any circulating RNA-binding proteins
• No phenol extractions, Proteinase K treatment nor carrier RNA required
• No time-consuming ultracentrifugation, filtration nor special syringes are required
• No precipitation reagents nor overnight incubation required
• Concentrate isolated exosomal RNA and free-circulating RNA into a flexible elution volume ranging from 50 µL to 100 µL
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different urine sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits since they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kits allow 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, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Urine Exosome and Free-Circulating RNA Isolation Mini Kit

For sample volumes ranging from 250 µL to 1 mL.

Urine Exosome and Free-Circulating RNA Isolation Midi Kit

For sample volumes ranging from 2 mL to 10 mL.

Urine Exosome and Free-Circulating RNA Isolation Maxi Kit

For sample volumes ranging from 11 mL to 30 mL.

Details

Supporting Data

Figure 1 / 6

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Kit Specifications
Minimum Urine Input	11 mL
Maximum Urine Input	30 mL
Size of RNA Purified	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 the specimen

*Please check page 5 of the product insert for the average yields and the common RNA quantification methods.

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipping. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.

Component	Cat. 59200 (50 preps)	Cat. 59300 (25 preps)	Cat. 59400 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	30 mL	50 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	2 x 20 mL	2 x 20 mL	2 x 20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	2 x 18 mL	18 mL	18 mL
Elution Solution A	2 x 6 mL	6 mL	6 mL
Mini Filter Spin Columns	50	25	15
Mini Spin Columns	100	50	30
Collection Tubes	100	50	30
Elution Tubes (1.7 mL)	100	50	30
Product Insert	1	1	1

• All in One. All the flexibility you need in a 5x Master Mix for multiplexing applications. NOW LYO READY!+ Sensitive. More free volume. 5x concentration allows more volume for target specific primers and probes (multiplexing).
  + Robust. Uniform amplification. Up to 30 target multiplexing in real-time (customer feedback). 
  + Fast TTR. No extraction needed. Reliable results with crude samples without extraction step, like blood.
  + Specific. No false amplification. Engineered Taq DNA polymerase more stable at room temperature. Aptamer-based hot-start prevents false amplification and provides a fast-start function.   
  + Lyo ready. Contains all necessary excipients needed for freeze-drying. Can be used for lyophilization. Enables RT storage and shipping once dried. Can be freeze-dried by us. Learn MoreFor research use and further manufacturing. Designed and manufactured under ISO13485.

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Exemplary tetraplex PCR assay

Detection of three specific pathogen targets and an internal control. Total input DNA per reaction was 10^6 (red), 10^5 (yellow), 10^4 (blue), 10^3 (green), 10^2 (purple), 10 (light blue) and 0 copies (grey) and 8000 copies/reaction for the internal control (shown in the CY5 plot).

5x Multiplex – robust PCR perforemance for a wide range of qPCR application

PlexTaq 5x qPCR Multiplex Master Mix contains all components necessary for rapid, sensitive and reproducible quantification of DNA and cDNA. An engineered DNA polymerase and an optimized buffer including ultrapure dNTPs are key components of the ready to use mix. A hot-start formulation of the included DNA polymerase prevents aptamer prevents false amplification and provides a fast start function. 

PlexTaq®´s formulation allows to use it also for direct PCRs from crude samples.

All in One. All the flexibility you need in a 5x Master Mix for multiplexing applications. NOW LYO READY!

+ Sensitive. More free volume. 5x concentration allows more volume for target specific primers and probes (multiplexing).
+ Robust. Uniform amplification. Up to 30 target multiplexing in real-time (customer feedback).
+ Fast TTR. No extraction needed. Reliable results with crude samples without extraction step, like blood.
+ Specific. No false amplification. Engineered Taq DNA polymerase more stable at room temperature. Aptamer-based hot-start prevents false amplification and provides a fast-start function.