This kit provides a fast, reliable and convenient method to isolate and concentrate exosomal RNA from previously purified exosomes. It allows for the isolation of RNA from intact extracellular vesicles (EVs) purified from different urine or plasma/serum sample volumes. The purification is based on Norgen’s proprietary resin.
The Exosomal RNA Isolation Kit is designed to isolate all sizes of RNA, including microRNA. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
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| Kit Specifications | |
| Sample type | Exosomes purified using Norgen’s Purification Kits and most other methods |
| Size of RNA Purified | All sizes, including miRNA and small RNA (< 200 nt) |
| Elution Volume | 50-100 μL |
| Time to Complete 10 Purifications | 35-40 minutes |
| Average Yields* | Variable depending on specimen |
*Please check page 5 of the product insert for the average yields and the common RNA quantification methods.
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
| Component | Cat. 58000 (50 preps) |
|---|---|
| Lysis Buffer A | 20 mL |
| Lysis Additive B | 2 mL |
| Wash Solution A | 18 mL |
| Elution Solution A | 6 mL |
| Mini Spin Columns | 50 |
| Collection Tubes | 50 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |
Description
ExcelTaq™ Klen-Taq DNA Polymerase is a specially blended enzyme mix containing KlenTaq-1 DNA polymerase (a 5’-exo-minus, N-terminal deletion of Taq DNA polymerase) and a small amount of a proofreading DNA polymerase. This unique blending helps to improve the fidelity, yield and processivity of the resultant PCR process. The Klen-Taq is also highly robust, showing high tolerance of varying concentrations of Mg2+; it is highly thermostable and has four times the fidelity compared to Taq DNA polymerase. The ExcelTaq™ Klen-Taq DNA Polymerase is ideal for DNA amplifications 0.5-5 kb in length on genomic DNA, and up to 10 kb on less complex templates.
Features
Storage
-20°C for 24 months
ExcelTaq™ Klen-Taq DNA Polymerase is a specially blended enzyme mix containing KlenTaq-1 DNA polymerase (a 5’-exo-minus, N-terminal deletion of Taq DNA polymerase) and a small amount of a proofreading DNA polymerase. This unique blending helps to improve the fidelity, yield and processivity of the resultant PCR process. The Klen-Taq is also highly robust, showing high tolerance of varying concentrations of Mg2+; it is highly thermostable and has four times the fidelity compared to Taq DNA polymerase. The ExcelTaq™ Klen-Taq DNA Polymerase is ideal for DNA amplifications 0.5-5 kb in length on genomic DNA, and up to 10 kb on less complex templates.
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.
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