96-Well PCR Plate / 0.2 mL Strip Chiller Block

Overview

• Isolate genomic DNA from all types of bacteria (both Gram-positive and Gram-negative)
• Rapid and convenient spin column protocol
• Purified bacterial gDNA has a minimal host gDNA contamination.
• High yield, high quality DNA for sensitive downstream applications including sequencing, PCR, qPCR and more

Synovial fluid is secreted into the joint cavity from the inner membrane of synovial joints. Synovial fluid is a plasma ultrafiltrate which contains proteins derived from both the blood plasma and produced by cells within the joint tissues. It lubricates the articulating joints as well as supplies oxygen and nutrients while removing carbon dioxide and metabolic wastes from the chondrocytes in the surrounding cartilage. Septic arthritis is usually caused by bacterial, viral or fungal infections to the synovial fluid. Staphylococcus aureus is the most common bacterial infection causing Septic arthritis. Diagnosing Septic arthritis is done through joint fluid (synovial fluid) analysis, blood tests or imaging tests.  

Norgen’s Synovial Fluid Bacterial Genomic DNA Isolation Kit provides a fast, reliable and simple procedure for isolating the highest quality and the highest quantity of bacterial genomic DNA from various amounts of synovial fluid ranging from 1 mL up to 10 mL.  Purification is based on using Norgen’s proprietary resin separation matrix. The kit is designed to isolate all Gram +ve and Gram -ve strains. Moreover, the kit allows the user to elute the purified bacterial genomic DNA into a flexible elution volume ranging from 25 µL to 50 µL. The purified bacterial gDNA is eluted in an Elution Buffer that is compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR, Southern Blot analysis and NGS.

Details

Supporting Data

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Kit Specifications
Maximum Input	2 x 109 bacterial cells
Column Binding Capacity	25 μg
Average Yield	up to 20 µg
Time to Complete 10 Purifications	45 minutes

Storage Conditions and Product Stability
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.

Component	Cat. 67900 (50 preps)
Precipitation Solution B	15 mL
Lysis Buffer T	6.5 mL
Solution BX	28 mL
Wash Solution A	38 mL
Elution Buffer B	8 mL
Micro Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

N-(Propargyl-PEG4-carbonyl)-N-bis(PEG1-methyl ester) is a crosslinker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage. The methyl ester groups can be hydrolyzed, reduced, or substituted under different conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

N-(Propargyl-PEG4-carbonyl)-N-bis(PEG1-methyl ester) is a crosslinker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage. The methyl ester groups can be hydrolyzed, reduced, or substituted under different conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Bioprocessing with Salt Active Nucleases  – High Salt Conditions

OverView

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.