This kit is suitable for high-throughput extraction of total RNA from paraffin embedded tissue samples, including miRNA. The kit is based on superparamagnetic magnetic particle purification technology. The extraction process does not require the use of toxic phenolic chloroform extraction or time-consuming alcohol precipitation.The whole extraction process only takes 120 minutes. The obtained RNA can be directly used in RT-PCR. The high-throughput extraction can be realized by 96 hole magnetic frame and 96 holeo scillator, various nucleic acid extractors (such as KingFisherFle, MagMix32, MagMix 96) or cooperating with the pipette workstation (Hamilton, Tican) for automatic extraction.
Detail
Introduction
This kit is suitable for high-throughput extraction of total RNA from paraffin embedded tissue samples, including miRNA. The kit is based on superparamagnetic magnetic particle purification technology. The extraction process does not require the use of toxic phenolic chloroform extraction or time-consuming alcohol precipitation.The whole extraction process only takes 120 minutes. The obtained RNA can be directly used in RT-PCR. The high-throughput extraction can be realized by 96 hole magnetic frame and 96 holeo scillator, various nucleic acid extractors (such as KingFisherFle, MagMix32, MagMix 96) or cooperating with the pipette workstation (Hamilton, Tican) for automatic extraction.
Details
Specifications
Features
Specifications
Main Functions
Isolation total RNA from FFPE tissue
Applications
RT-PCR, cDNA synthesis, second generation sequencing
Products
RNA, miRNA
Purification method
Polydisperse magnetic beads
Purification technology
Magnetic beads technology
Process method
Manual or automatic
Adaptive instrument
Nucleic acid extractor, pipetting workstation
Sample type
FFPE slice, FFPE embedded tissue
Sample amount
≤6 slices
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
Fast – several samples can be extracted in 120 minutes by column method
Safe – no phenol chloroform extraction required
Kit Contents
Contents
IVD3022
Purification Times
200 Preps
MagBind Particles
4.5 ml
Proteinase K
100 mg
Protease Dissolve Buffer
6 ml
DNase I
4 x 600 µl
DNase Buffer
30 ml
Buffer DPS
200 ml
Buffer FRL
40 ml
Buffer AL
40 ml
Buffer MW1*
110 ml
Buffer MW2*
2 x 50 ml
Nuclease Free Water
30 ml
Storage and Stability
Proteinase K and MagBind Particles should be stored at 2–8°C upon arrival. DNase I should bestored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K and MagBind Particles up to 8 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 theseconditions.
Other Products
Bis-propargyl-PEG13
Product Info
Document
Product Info
Bis-propargyl-PEG13 comprises two propargyl groups which can form triazole linkages with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry reactions. The hydrophilic PEG units help improve the solubility of the molecule in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
Bis-propargyl-PEG13 comprises two propargyl groups which can form triazole linkages with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry reactions. The hydrophilic PEG units help improve the solubility of the molecule in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Bioprocessing with Salt Active Nucleases – High Salt Conditions
Product Info
Document
Product Info
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.
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.
The 16S V2-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V2-V3 amplicon libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XP Magnetic Beads (supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.
The 16S V2-V3 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the V2-V3 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XP beads. The libraries are then ready for quantification, pooling and sequencing.
Storage Conditions and Product Stability Norgen’s 16S V2-V3 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.
All kit components should remain stable for at least 1 year when stored at the specified storage conditions.
