The NGS Single Stranded DNA Library Prep Kit (illumina platform) was developed for construction of high quality libraries using sheared single stranded DNA (50 ng – 500 ng) as input. The kit is ideal for making NGS libraries with samples of denatured DNA, viral DNA, highly degraded ancient DNA and other single stranded DNA. The workflow of the ssDNA kit is simple: make the libraries in two steps followed by PCR and cleanup steps. The libraries will be ready in just 1.5 hours with a 10 minutes of hands-on time. The library pooling is possible dependent on the type of index. The final library is strand specific.
Detail
The NGS Single Stranded DNA Library Prep Kit (illumina platform) was developed for construction of high quality libraries using sheared single stranded DNA (50 ng – 500 ng) as input. The kit is ideal for making NGS libraries with samples of denatured DNA, viral DNA, highly degraded ancient DNA and other single stranded DNA. The workflow of the ssDNA kit is simple: make the libraries in two steps followed by PCR and cleanup steps. The libraries will be ready in just 1.5 hours with a 10 minutes of hands-on time. The library pooling is possible dependent on the type of index. The final library is strand specific.
NGS Single Stranded DNA Library Prep Kit workflow
Three index types are available for the kit:
Non-index (Cat.# 30081): Libraries do not have index.
Index(Cat.# 30082): Each of the index primers has a unique 6-base index sequence that can be used to identify libraries. ssDNA library multiplexing is up to 48 samples. Index information can be downloaded here.
Unique dual index (Cat.# 30083): ssDNA sample multiplexing up to 96 libraries is possible with unique dual indexes. We have developed a Four-Base Difference Index System. This makes it possible to generate indexes with at least 4 bases different from each other in the 8 bases index length. The unique dual indexing primers remove NGS errors (example: de-multiplexing errors, read mis-assignment, index hopping etc). Index information can be downloaded here.
Kit advantages:
Quick and simple Protocol
Quick: Total protocol time is only 1.5 hours
Simple: Hands-on time only 10 minutes
Easy procedure based on:
The ready-to-use master mix: Made reaction setup easy
Less reaction components: Simplify the reaction preparation
Less magnetic beads needed: Reduced more than 50% of the beads for cleanup steps
Directional library
Single stranded DNA as input: From 50 ng to 500 ng
NGS Single Stranded DNA Library Prep Kit has similar library conversion efficiency and yield as compared to a regular DNA library prep kit.
Alignment rate and duplication rate: comparison of single stranded DNA library kit versus double stranded DNA library prep kit.
Other Products
bis-PEG23-endo-BCN
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Product Info
Bis-PEG23-endo-BCN is a click chemistry reagent, The BCN group enable copper free click chemitry with azide-tagged molecules. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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Bis-PEG23-endo-BCN is a click chemistry reagent, The BCN group enable copper free click chemitry with azide-tagged molecules. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Bioprocessing with Salt Active Nucleases – High Salt Conditions
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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.
DBCO-PEG8-amine TFA salt is a PEG linker which contains DBCO and amine moieties. The DBCO group is commonly used in copper-free Click Chemistry reactions. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The hydrophilic PEG spacer increases the water solubility of the compound. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
DBCO-PEG8-amine TFA salt is a PEG linker which contains DBCO and amine moieties. The DBCO group is commonly used in copper-free Click Chemistry reactions. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The hydrophilic PEG spacer increases the water solubility of the compound. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
