These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating DNA, circulating RNA including exosomal RNA as well as viral DNA/RNA from fresh, frozen or preserved urine samples. Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix. These kits are designed to isolate all sizes of cfc-DNA and cfc-RNA, including microRNA, as well as all sizes of exosomal RNA. Norgen’s Urine Cell-Free Circulating and Viral Nucleic Acid Purification Kits provide a clear advantage over other available kits in that they do not require phenol/chloroform or any protease treatments. Moreover, the kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The purified nucleic Acid is of the highest integrity, and can be used in a any downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, methylation-sensitive PCR and Southern Blot analysis, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.
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Background
Recent evidence indicates that cell-free circulating (cfc) nucleic acids such as DNA and RNA, including exosomal RNA, in urine contains valuable information for the discovery of biomarkers that can help with early detection of certain cancer types, for monitoring the disease status as well as for the detection fetal DNA. Moreover, urine has been widely used for the detection of viral infections, including HIV, HCV and HBV.The advantage for using urine as a source for cancer biomarkers is that it can be acquired in large quantities without using invasive procedures, and repeated sampling from the same individual is applicable, which facilitates longitudinal studies. There are many advantages favouring the use of urinary nucleic acids 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 acids is similar to that in urine or serum; (3) nucleic acid purification from urine is technically much easier because of its low protein concentration (-fold lower than blood).
Urine Cell-Free Circulating and Viral Nucleic Acid Purification Mini Kit
For sample input volumes ranging from 250 µL to 2 mL.
Urine Cell-Free Circulating and Viral Nucleic Acid Purification Midi Kit
For sample input volumes ranging from 2 mL - 10 mL.
Urine Cell-Free Circulating and Viral Nucleic Acid Purification Maxi Kit
For sample input volumes ranging from 10 mL - 30 mL.
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Paramagnetic beads are added to the sample, and their surfaces attract nucleic acids to bind to the beads. Using a strong magnet, the beads are held in place while removing unwanted material. After washing, the genetic material is eluted from the beads in water or a low-salt buffer. Using Dynabeads magnetic beads, many separation applications can be achieved.
Magnetic bead isolation is now one of the most popular nucleic acid extraction methods due to its scalability and automation compatibility. MagMAX bead kits and KingFisher Sample Purification Systems are designed to work together to efficiently purify a variety of nucleic acids.
Nucleic acid isolation by organic extraction involves the addition of phenol and guanidine isothiocyanate to separate the DNA, RNA, and proteins into different organic phases. Organic extraction is a low-cost method, and with advanced reagents such as TRIzol or phenol-chloroform, are straightforward processes requiring less lab equipment.
After lysis is performed and the solvents are added, the mixture then goes through a process of centrifugation to separate the aqueous phase (containing nucleic acids) from the organic phase (containing proteins and lipids).
Nucleic acids bind to silica (also known as glass fibers) under high-salt conditions and can be released under low-salt conditions. Silica-containing columns provide an easy way to bind, wash, and elute purified nucleic acids from multiple clarified cell lysates in parallel.
Invitrogen Purelink and GeneJET columns are designed to flow buffers through centrifugation, vacuum, or gravity. Most protocols use spin column technology to take advantage of readily available lab equipment. Spin plates provide a high-throughput format based on the same isolation principle.
The KingFisher PlasmidPro Maxi Processor leverages the same processes and chemistry as traditional spin column-based plasmid purification methods with the advantages of automation. With prefilled reagents in a single cartridge, it helps eliminate the need for setup, centrifugation, and pipetting, reducing errors in sample preparation. This fully automated system with its "press and go" design is the only one of its kind for maxi-scale plasmid DNA purification.
Thermo Fisher Scientific offers a wide range of plasmid DNA extraction products that are designed to isolate plasmid DNA at the purity and scale you need and to support column-based workflows. Choose the PureLink or GeneJet kits based on purity grade and culture volume. Use our selection tool to find the right plasmid purification kit for your research.