N-(Propargyl-PEG2)-N-bis(PEG3-t-butyl ester) is a 3-arm PEG reagent with a propargyl groups and two t-butyl ester groups. The propargyl group can react with azides via copper catalyzed Click Chemistry reaction to yield a stable triazole linkage. The t-butyl ester groups can be deprotected under acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
N-(Propargyl-PEG2)-N-bis(PEG3-t-butyl ester) is a 3-arm PEG reagent with a propargyl groups and two t-butyl ester groups. The propargyl group can react with azides via copper catalyzed Click Chemistry reaction to yield a stable triazole linkage. The t-butyl ester groups can be deprotected under acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Bioprocessing with Salt Active Nucleases – High Salt Conditions
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.
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.
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).
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.
This kit provides a rapid method for the isolation and purification of small RNA molecules (< 200 nt) from cultured animal cells, small tissue samples, bacterial cells, plants, and blood.
Two columns are provided with this kit. The first column captures large RNA, while the small RNA are captured on a second column and are eluted concentrated in 25 µL of nuclease-free water.
The small RNA can be used in various downstream applications relating to miRNA profiling, gene regulation, and functional analysis. The eluted RNA is ready for RT-qPCR, microarrays, and NGS applications.
Background
These small RNAs include regulatory RNA molecules such as microRNA (miRNA) and short interfering RNA (siRNA), as well as tRNA and 5S rRNA. Small RNA molecules are often studied due to their ability to regulate gene expression. miRNAs and siRNAs are typically 20-25 nucleotides long and regulate gene expression by binding to mRNA molecules and affecting their stability or translation.
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| Kit Specifications | |
| Maximum Column Binding Capacity | Up to 50 μg RNA |
| Maximum Column Loading Volume | 650 μL |
| Minimum Elution Volume | 20 μL |
| Size of RNA Purified | All sizes, including small RNA < 200 nt |
| Time to Complete 10 Purifications | 25 minutes |
| Maximum Amount of Starting Material: Animal Cells Animal Tissues Bacteria Plant Tissues Blood | 3 x 106 cells 5-25 mg 1 x 109 cells 50 mg 100 μL |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 21300 (25 preps) |
|---|---|
| Buffer RL | 40 mL |
| Wash Solution A | 38 mL |
| Elution Solution A | 6 mL |
| Large RNA Removal Column | 25 |
| microRNA Enrichment Column | 25 |
| Collection Tube | 50 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |
Fig 1. Only three steps will be need for Sample Preparation and only 20 minitutes will be taken for Sample Preparation.
Fig 2. Seven concentration samples of 0.3fg/μL, 3fg/μL, 30fg/μL, 300fg/μL, 3pg/μL, 30pg/μL, 300pg/μL were detected. CV of each concentration was < 30%, Regression coefficient associated with standard solutions was 0.99992, and amplification efficiency was 100.370%.
Fig 3. Five concentration samples of 0.1fg/μL, 0.3fg/μL, 0.5fg/μL, 1fg/μL and 3fg/μL were detected, and 10 multiple wells were detected for each concentration. The CV of concentration values of samples with 0.3fg/μL and above concentrations were less than 30%.
Fig 4. DNA recovery can be determined by including samples spiked with known DNA amounts which are prepared from the corresponding DNA standards. Typically, the range for this value varies from 70% to 130%.
Fig 5. Only one Reagent for qPCR MIX.
Note: Price not include shipment & duty, contact us to get full quote.
The resDNASEQ CHO Residual DNA Quantitation kit is designed for the quantification of residual DNA from CHO, in cell lines which are used for production of biopharmaceutical products. The Ducky Bio residual DNA CHO Assay, based on proven real-time qPCR technology, makes testing of residual DNA from the Chinese hamster ovary (CHO) cell line rapid, specifc. The PCR-based assay is sensitive and specific for DNA from the CHO cell line and not subject to detection of human or environmental DNA that might be introduced during sample handling. The kit was developed to meet the sensitivity requirements defined by WHO (10 ng CHO DNA per therapeutic dose).
