Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.
The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.
Packaged, optimised and ready to use. Expect Better Data.
Primer and probe mix (150 reactions)
Reverse Transcription, target specific primers (RNA genome viruses only)
Copy number standard curve (sufficient for multiple standard curves)
Internal extraction control – Read through VIC channel*
Endogenous control (150 tests)
RNAse/DNAse free water
*alternative fluorophores available on request
These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different plasma/serum sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA as well as all sizes of the free-circulating protein-bound RNA, including microRNA. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, these kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA 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.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit
For sample volumes ranging from 50 µL to 1 mL.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Midi Kit
For sample volumes ranging from 1 mL to 4 mL.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit
For sample volumes ranging from 4 mL to 10 mL.
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Kit Specifications | |
Minimum Plasma Input | 50 μL |
Maximum Plasma Input | 1 mL |
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 solutions 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.
Important Note
This kit is suitable for the purification of exosomes from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.
Component | Cat. 59500 (50 preps) | Cat. 59600 (25 preps) | Cat. 59700 (15 preps) |
---|---|---|---|
Slurry E | 12.5 mL | 12.5 mL | 12.5 mL |
ExoC Buffer | 8 mL | 8 mL | 2 x 8 mL |
ExoR Buffer | 12 mL | 12 mL | 12 mL |
Lysis Buffer A | 2 x 20 mL | 2 x 20 mL | 2 x 20 mL |
Lysis Additive B | 2 mL | 2 mL | 2 mL |
Wash Solution A | 2 x 18 mL | 18 mL | 18 mL |
Elution Solution A | 2 x 6 mL | 6 mL | 6 mL |
Mini Filter Spin Columns | 50 | 25 | 15 |
Mini Spin Columns | 100 | 50 | 30 |
Collection Tubes | 100 | 50 | 30 |
Elution Tubes (1.7 mL) | 100 | 50 | 30 |
Product Insert | 1 | 1 | 1 |
Different-sized amplicons from 3 ng of a DNA plasmid were amplified. The use of Taq DNA polymerase resulted in clean and high yield of products, as analysed after PCR on a 0.8% agarose gel.
A fragment (64 bp) of the human blood-coagulation factor IIa (F2) was amplified from 20 ng, 2 ng, 200 pg and 20 pg of a human genomic DNA extract. The same experiment was performed in parallel using the Taq DNA polymerase mix from another well-established and known supplier. PCR products were subsequently analysed on a 2.5% agarose gel.
Non-specific amplification is prevented by using this Taq DNA polymerase variant as the DNA polymerase is hotstart-formulated. The 10x reaction buffer supplied together with the DNA polymerase has been specifically designed for optimal PCR performance and DNA polymerase activity. This allows the use of this DNA polymerase in a wide range of PCR applications.
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