Introduction

This product is suitable for rapid extraction of DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from blood, buffy coat, tissue and other samples
Applications	Second generation sequencing, PCR, real time PCR, etc.
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Anticoagulant blood, concentrated blood, buffy coat, lymphocytes and cultured cells
Sample amount	Whole blood :< 200μl; Saliva / swab:< 400μl; Tissue :< 20mg
Yield	0.1 – 50μg
Elution volume
Time per run

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and protease. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• High purity – OD 260 / 280 : 1.7- 1.9, OD 260 / 230 : 1.5 – 2.0
• Economy – less than 50% of the price of Qiagen and other imported products
• High yield – most optimal process, ensuring the recovery up to 90%
• Strong processing ability – samples including animal blood, cultured cells, animal tissues, etc.

Kit Contents

Contents	IVD3102
Purification Times	200
MagPure Particles	5 ml
Proteinase K	100 mg
Protease Dissolve Buffer	10 ml
Rnase A	40 mg
Buffer ATL	60 ml
Buffer AL	60 ml
Buffer BD*	20 ml
Buffer BW1*	110 ml
Elution Buffer	30 ml

Cat.No	Reagent	IVD3102-F-96
Proteinase K		50 mg
Protease Dissolve Buffer		6 ml
RNase A		20 mg
Buffer ATL		30 ml
Buffer AL		30 ml
96-Tip		1
Sample plate (DW Plate)	450µl Buffer BD(Ethanol Added)	1
Wash 1 Plate (DW Plate)	600µl Buffer BW1(Ethanol Added)	1
Wash 2 Plate (DW Plate)	600µl Buffer BW1(Ethanol Added)	1
Wash 3 Plate (DW Plate)	750µl Buffer GW2, 20µl MagPure Particle	1
Elution plate (DW Plate)	80µl Elution Buffer	1

Cat.No	Reagent	IVD3102-TL-06
Proteinase K		50 mg
RNase A		20 mg
Protease Dissolve Buffer		6 ml
Buffer ATL		40 ml
Buffer AL		40 ml
AS-Tip		12
2.0ml V-bottom plate	Row 1/7：450µl Buffer BDRow 2/8：450µl Buffer BW1Row 3/9：450µl Buffer BW1Row 4/10：20μl Magpure Particle450μl Wash Buffer GW2 Row 5/11：450μl Wash Buffer GW2 Row 6/12：80µl Elution Buffer	6

Storage and Stability

Proteinase K, RNase A, MagPure Particles should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

Experiment Data

This product is suitable for rapid extraction of DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.

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.