Propargyl-PEG13-bromide is a heterobifunctional reagent that can participate in copper catalyzed azide-alkyne Click Chemistry to form a stable triazole linkage. The bromide (Br) can be used in nucleophilic substitution reactions. The PEG units increase water-solubility of the molecule in in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Detail
Propargyl-PEG13-bromide is a heterobifunctional reagent that can participate in copper catalyzed azide-alkyne Click Chemistry to form a stable triazole linkage. The bromide (Br) can be used in nucleophilic substitution reactions. The PEG units increase water-solubility of the molecule in in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Hipure Microbial RNA kit is suitable for extracting high-purity total RNA from bacterial/yeast culture medium. This kit combines high-efficiency Magzol Reagent (one-step extraction reagent) and silicagel column purification technology to complete the extraction of high-purity total RNA in only 40 minutes. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.
Details
Specifications
Features
Specifications
Main Functions
Isolation total RNA from bacteria, yeast cells
Applications
RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Bacteria, yeast cells
Sample amount
Bacteria: <10^9;Yeast cells:<2 x10^7
Elution volume
≥30μl
Time per run
≤40 minutes
Liquid carrying volume per column
800µl
Binding yield of column
100µg
Principle
Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such as guanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein and other impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.
Advantages
Fast – several samples can be extracted in 30 minutes
High purity – the purified RNA can be directly used in various downstream applications
High recovery – RNA can be recovered at the level of PG
Good repeatability – silica gel column purification technology can obtain ideal results every time
Broad spectrum – it can deal with various bacteria, including Gram-positive bacteria that are difficult to be lysed
Sufficient components – the kit contains carried wall breaking enzymes and glass beads
Kit Contents
Contents
R418202
R418203
Purification Times
50 Preps
250 Preps
HiPure RNA Mini Columns
50
250
2ml Collection Tubes
50
250
Glass Beads (0.1-0.6mm)
30 g
150 g
DNase I
600 μl
5 x 600 μl
DNase Buffer
6 ml
30 ml
Protease Dissolve Buffer
1.8 ml
15 ml
Buffer ATL
50 ml
200 ml
Buffer PHC
50 ml
200 ml
Buffer GXP2*
20 ml
100 ml
Buffer RW1
50 ml
250 ml
Buffer RW2*
20 ml
2 x 50 ml
RNase Free Water
10 ml
30 ml
Storage and Stability
Buffer PHC should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Buffer PHC up to 12 weeks) at room temperature(15–25°C) does not affect their performance. The remaining kit components can be stored at roomtemperature (15–25°C) and are stable for at least 18 months under these conditions.
Document
Hipure Microbial RNA kit is suitable for extracting high-purity total RNA from bacterial/yeast culture medium. This kit combines high-efficiency Magzol Reagent (one-step extraction reagent) and silicagel column purification technology to complete the extraction of high-purity total RNA in only 40 minutes. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.
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.