Influenza virus infection of birds, humans and other animals is a major public health problem worldwide. Influenza viruses are classified as either type A, B or C based on differences in their nucleoproteins and matrix proteins. The type A viruses are the most virulent human pathogens among the three influenza types and cause the most severe disease and epidemics. The different types can be further classified into subtypes based on antigenic differences in two surface glycoproteins; hemagglutinin and neuroamidase. All known subtypes of influenza A can be found in birds (H1-H16, N1-N9), while a limited number of the subtypes have been found in humans (H1-H3, N1 and N2). However, over the past few years, various subtypes of Influenza A viruses, including H5N1, have been reported to infect humans (WHO, 2006). In addition, the coexistence of human influenza viruses and avian influenza viruses may provide an opportunity for genetic material to be exchanged between these viruses. This could potentially create a new virulent influenza strain that is easily transmissible and lethal to humans (Food Safety Research Information Office, 2006). Thus, there is the need for sensitive diagnostic tests to allow for the rapid and early detection of these H5 influenza virus infections, to help reduce the risk of epidemics or pandemics in both animals and humans.
H5N1 TaqMan RT-PCR Kit, 100 reactions
H5N1 TaqMan RT-PCR Probe/Primer Set and Controls, 100 reactions
For research use only and NOT intended for in vitro diagnostics.
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Storage Conditions and Product Stability
All kit components can be stored for 1 year after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
| Component | Cat. TM35450 (100 preps) | Cat. TM35410 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| H5N1 Primer & Probe Mix | 280 μL | 280 μL |
| H5N1 Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating RNA, including exosomal RNA as well as viral RNA from fresh, preserved or frozen urine samples. All components for the purification are provided in one convenient and fast kit for the easy processing of small input volumes of bodily fluids. The purified urine RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.
Background
Recent evidence indicates that cell-free circulating RNA (cf-RNA) including exosomal RNA in urine contains valuable information for the discovery of biomarkers that can help for the early detection of certain cancer types and for monitoring the disease status as well as for the detection of any infectious pathogens. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. The advantage for using urine as a source for cancer biomarkers is that it can be acquired in large quantities without using invasive procedures. In addition, repeated sampling from the same individual is applicable, which facilitates longitudinal studies. There are many advantages favouring the use of urinary nucleic acid for cancer biomarker discovery over blood, tissue samples or other bodily fluids, including: (1) urine is non-infectious for HIV and less infectious for many other pathogens; (2) the profile of urinary nucleic acid is similar to that in plasma or serum but with a lower concentration; (3) Nucleic acid purification from urine is technically much easier because of its low protein concentration (1000-fold lower than blood).
Urine Cell-Free Circulating RNA Purification Mini Kit
For sample volumes ranging from 250 µL to 2 mL.
Urine Cell-Free Circulating RNA Purification Midi Kit
For sample volumes ranging from 2 mL to 10 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.
Urine Cell-Free Circulating RNA Purification Maxi Kit
For sample volumes ranging from 10 mL to 30 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL
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| Kit Specifications | |
| Minimum Urine Input | 10 mL |
| Maximum Urine Input | 30 mL |
| Size of RNA Purified | All sizes including miRNA and small RNA (< 200 nt) |
| Elution Volume | 50-100 μL |
| Time to Complete 10 Purifications | 40-45 minutes |
| Average Yields | Variable depending on specimen |
*Please check page 6 of the product insert for average yields and common RNA quantification methods.
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. These kits are 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. 56900 (50 preps) | Cat. 57000 (20 preps) | Cat. 57100 (10 preps) |
|---|---|---|---|
| Binding Solution K | 25 mL | 75 mL | 1 x 75 mL 1 x 25 mL |
| Lysis Buffer A | 30 mL | 20 mL | 20 mL |
| Wash Solution A | 18 mL | 18 mL | 18 mL |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Mini Spin Columns | 50 | 20 | 10 |
| Midi Spin Columns | – | 20 | – |
| Maxi Spin Columns | – | – | 10 |
| Collection Tubes | 50 | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 |
Propargyl-PEG3-alcohol consists of a hydroxyl group and a propargyl group. The propargyl group reacts with azide compounds in copper catalyzed azide-alkyne Click Chemistry to form a triazole linkage. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Propargyl-PEG3-alcohol consists of a hydroxyl group and a propargyl group. The propargyl group reacts with azide compounds in copper catalyzed azide-alkyne Click Chemistry to form a triazole linkage. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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