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Norgen’s Urine (Exfoliated Cell) DNA Isolation Kit is designed for the rapid preparation of genomic DNA from exfoliated cells that have been shed into the urine from the urinary tract. The genomic DNA isolated from these exfoliated cells can be used in a number of diagnostic and research applications including the diagnosis and monitoring of bladder,kidney, or other urinary tract cancers. The kit allows for the isolation of genomic DNA from exfoliated cells present in 1 to 50 mL of urine. The kit preferentially purifies DNA from other cellular proteinaceous components, as well as from the contaminating species found in urine such as glucose and salts. Typical yields of DNA will vary depending on the cell density of the urine sample, which is affected by a number of factors including health, diet and sex of the individual donating the urine. The purified DNA is of the highest quality and can be used in a number of downstream applications.
Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. Briefly, the exfoliated cells in the urine are pelleted, then Resuspension Solution and Proteinase K are added to the pellet. Next, the cells are lysed using the provided Lysis Solution. Binding Solution is then added to the lysate, and the solution is loaded onto a spin column (BIND). Under these conditions on the DNA will bind to Norgen’s resin while most of the RNA and proteins will be removed in the flowthrough. The bound DNA is then washed in order to remove any remaining impurities (WASH). Lastly, the purified genomic DNA is eluted into the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart to the right.
FEATURES AND BENEFITS
Fast processing -Rapid spin-column format allows for the processing of multiple samples in 45 minutes (inlcuding a 30 minute incubation).
Isolate genomic DNA from small volumes of urine - DNA can be isolated and detected from the exfoliated cells found in as little as 1 mL of urine.
High quality DNA - Removal of highly concentrated salts, metabolic wastes and proteins provides high quality DNA to be used in various downstream applications.
Recovered DNA is suitable for downstream applications - Purified DNA is fully compatible with PCR, quantitative PCR and Southern Blot analysis.
APPLICATIONS
The purified genomic DNA is compatible with downstream applications such as:
Figure 1: Isolation and Detection of Biologically Active Genomic DNA from 1 mL of Urine. Genomic DNA was isolated from the exfoliated cells found in a 1 mL urine sample using Norgen’s Urine (Exfoliated Cell) DNA Isolation Kit. The purified DNA was eluted in 100 µL of Elution Buffer, and 5 µL of the isolated DNA was used as a template in quantitative PCR using human beta actin gene primers to detect the genomic DNA using the iQ SYBR Green Supermix (BioRad, #170-8882). The red lines in the PCR baseline graph above correspond to DNA standards, while the blue line corresponds to the successful PCR results when DNA isolated from the exfoliated cells in 1 mL of urine was used as the template.
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. All reagents should remain stable for at least 1 year in their unopened containers.
PROTOCOL
Urine (Exfoliated Cell) DNA Purification Kit - Protocol PDF version
PRODUCT INFORMATION SHEETS
Urine (Exfoliated Cell) DNA Purification Kit - Product Information Sheet PDF version
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