Urine DNA Isolation Kits Fast and reliable purification of genomic and apoptotic DNA from urine For research use only and NOT intended for in vitro diagnostics. CE-IVDD marked diagnostic slurry format available here Powered by Bioz Urine DNA Isolation Kits Fast and reliable purification of genomic and apoptotic DNA from urine SKU 18100 Format Spin Column Slurry Maxi Slurry Size 50 Preps Contact Distributor Overview Details Documentation Citations Overview Rapid isolation of both small and large species of DNA from urine Convenient spin column format Effective removal of PCR inhibitors Purified DNA is highly suited to sensitive downstream applications Allows for the purification of viral DNA from urine Both high molecular weight DNA (greater than 1 kb in size; mostly cell associated) and the smaller DNA (150 - 250 bp; derived from the circulation) is effectively isolated and purified using a rapid and convenient spin column protocol. This kit can be used to isolate DNA from a broad range of viruses in urine as well. Salts, metabolic wastes, proteins and other contaminants are removed to yield inhibitor-free DNA for use in sensitive applications. The DNA is of excellent quality for various downstream applications such as PCR, qPCR and DNA fingerprinting, methylation studies and more. This kit is fully compatible with Norgen's Urine Collection and Preservation Tubes. Urine DNA Isolation Kit This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 50 μL to 1.75 mL of urine. Multiple samples can be processed in 30 minutes. Urine DNA Isolation Kit (Slurry Format) This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 3 mL to 25 mL. Multiple samples can be processed in 30 minutes. Multiple samples can be processed in 45 minutes. Urine DNA Isolation Maxi Kit (Slurry Format) This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 25 mL of urine up to 80 mL. Multiple samples can be processed in 45 minutes. Background DNA found in urine can be divided into 2 basic categories. The larger species, genomic-DNA (gDNA), is generally greater than 1 kb in size, and appears to be derived mainly from exfoliated cells. The second species is smaller, generally between 150 and 250 bp (apoptotic-DNA), and derives, at least in part, from the circulation. The second species is also considered as an RNA/DNA hybrid as reported by Halicka et al. (2000). Both types of DNA can be isolated reliably using this kit. Details Supporting Data Previous Figure 1. A Typical Agarose Gel Showing Total Urinary DNA Isolated from 1.5 mL of Urine using Norgen's Urine DNA Isolation Kit. Total urinary DNA was isolated from two different 1.5 mL urine samples. The samples were processed according to the bind, wash and elute procedure provided in the kit, and the DNA was eluted into 2 separate elution volumes. The proteins were first eluted into 100 µL of Elution Buffer (E1), followed by a second elution using 75 µL of Elution Buffer (E2). The purified urine DNA was then run on a 1.2% agarose gel, and each ane contains one tenth from each elution (i.e. E1: 10 µL out of 100 µL were loaded on the gel, E2: 7.5 µL out of 75 µL were loaded on the gel ). Lane M is 10 µL of Norgen’s FastRunner DNA Ladder. Both the large and small urine DNA species can be seen on the gel. Figure 2. Isolation and Detection of DNA from 1.75 mL Urine Samples. Total genomic DNA was isolated from two different 1.75 mL urine samples using Norgen’s Urine DNA Isolation Kit. The bind, wash and elute procedure was performed, and the purified DNA was eluted into two separate elutions of 100 µL (E1) and 75 µL (E2). The isolated DNA was then subjected to quantitative PCR using human 5S gene primers to detect the genomic DNA using the iQ SYBR Green Supermix (BioRad, #170-8882). Five microliters from each elution was used as a template in a 20 µL qPCR reaction. The red line in the PCR baseline graph above corresponds to the first elution from the DNA isolated from the first urine sample, the green line corresponds to the second elution from the DNA isolated from first urine sample, the blue line corresponds to the first elution from the DNA isolated from the second urine sample, whereas the orange line corresponds to the second elution from the DNA isolated from the second urine sample. The yellow line corresponds to the No Template Control. Figure 3. Circulating DNA Isolated from Urine can be used as the Template in PCR Reactions.Total urinary DNA was isolated from three different 1.5 mL urine samples using Norgen's Urine DNA Isolation Kit. The bind, wash and elute procedure was performed, and the purified DNA was eluted into two separate elutions of 100 µL (E1) and 75 µL (E2). Five microliters of each elution was then used as a template in a PCR reaction to amplify the K-ras gene. Lanes A-C contain the expected 157 bp product, and correspond to the first elution from each sample. Lane D is the positive control of 293 HEK DNA and shows the expected 157 bp product, while Lane E is the negative control. Lane M is Norgen's FastRunner DNA Ladder. Figure 4. Typical Agarose Gel Showing Total Urinary DNA Isolated from Different Urine Volumes using Norgen's Urine DNA Isolation Kit (Slurry Format). Total urinary DNA was isolated from 3 mL, 6 mL, 12 mL and 24 mL of urine. Total urinary DNA was isolated from each urine sample according to the isolation protocol that is optimized for different sample volumes. The isolated DNA was eluted into two separate elutions (E1 and E2). The purified urine DNA was then loaded onto a 1.5% agarose gel. Each lane shows one tenth from each elution. It can be seen that the first elution contains most of urinary DNA whereas the second elution contains the rest of the urinary DNA isolated. Lane M is 10 µL of Norgen's FastRunner DNA Ladder. Figure 5. Isolation and Detection of DNA from Different Volumes of Urine. Total genomic DNA was isolated from 3, 6, 12 and 24 mL urine samples using Norgen's Urine DNA Isolation Kit (Slurry Format). The urinary DNA was isolated from each urine sample according to the provided protocol, and the DNA was eluted into two separate elutions (E1 and E2). The isolated DNA was then subjected to quantitative PCR using human 5S gene primers to detect the genomic DNA. To test the absence of PCR inhibitors usually accompanying DNA isolated from urine, an increasing amount from each elution (1, 3, 6 and 9 µL) was used as a template in the PCR reaction. Urinary genomic DNA was successfully detected from all the different urine volumes with no sign of PCR inhibition, even when 9 µL of urine DNA was used as the template. Figure 6. Highly Sensitive Isolation of Viral DNA from 25 mL of Urine. Herpes Simplex Virus - 1 (HSV-1) sequences were cloned into a plasmid, and 25 mL urine samples were then spiked with decreasing amounts of the plasmid (106, 105, 104, 103, 102 and 101). Total DNA was then isolated using Norgen’s Urine DNA Isolation Kit (Slurry Format). Ten microlitres of each 100 µL elution was then used as the template in a 20 µL PCR reaction using HSV-1-specific primers and Norgen's 2X PCR Master Mix (Cat #28007). Each PCR reaction was then run on a 1.5% agarose gel for visual analysis. The HSV-1 amplification product is 275 bp. As few as 250 HSV-1 viral copies could be isolated and detected from 25 mL of urine. Figure 7. Typical Agarose Gel Showing Total Urinary DNA Isolated from Different Urine Volumes using Norgen's Urine DNA Isolation Maxi Kit (Slurry Format). Total urinary DNA was isolated from 20 mL, 40 mL, 60 mL and 80 mL of urine. Total urinary DNA was isolated from each urine sample according to the isolation protocol that is optimized for different sample volumes. The isolated DNA was eluted, and was then loaded onto a 1.8% agarose gel. Each lane shows one tenth from each elution. It can be seen that urine DNA is increasing linearly with the urine input volume. It should also be noted that the circulating DNA started to appear in the form of ladder with the increase of the urine sample input. Lane M is 10 µL of Norgen's FastRunner DNA Ladder. Figure 8. Linearity of Ct Values and DNA Yields When Scaling Up Urine Input. Total urinary DNA was isolated from 20 mL, 40 mL, 60 mL and 80 mL of urine. Total urinary DNA was isolated from each urine sample according to the isolation protocol that is optimized for different sample volumes. The isolated DNA was then subjected to quantitative PCR using human 5S gene primers to detect the genomic DNA. Urinary genomic DNA was successfully detected from all the different urine volumes with no sign of PCR inhibition and in a linear relationship to the total urine DNA yield (Ct values decrease with increasing the sample urine input) Next Click for expanded view Kit Specifications Volume of Urine Processed 1.75 mL Average Yield* Up to 50 ng Size of DNA Purified Large (> 1 kb) and small (150-250 bp) Time to Complete 10 Purifications 30 minutes hands-on time (plus a 1 hour incubation) * Yield will vary depending on the type of sample processed 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. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature. Component Cat. 18100 (50 preps) Cat. 48800 (50 preps) Cat. 50100 (50 preps) Binding Solution K 15 mL - - Slurry B1 - 18 mL 18 mL Proteinase K in Storage Buffer 2 mL - - Pronase K in Storage Buffer 2 mL - - Soluton WN 9 mL - - Binding Buffer A - - 50 mL Lysis Buffer A - 30 mL 30 mL Wash Solution A - 38 mL 38 mL Wash Solution B 30 mL - - Wash Solution D 9 mL - - Binding Solution K 15 mL - - Elution Buffer B 15 mL 15 mL 15 mL Micro Spin Columns 50 - - Mini Filter Spin Columns - 50 50 Collection Tubes 50 50 50 Elution Tubes (1.7 mL) 100 100 100 Product Insert 1 1 1 Documentation PROTOCOLS (18100) Urine DNA Isolation Kit - Protocol (50 prep) (48800) Urine DNA Isolation Kit (Slurry Format) - Protocol (50 prep) (50100) Urine DNA Isolation Maxi Kit (Slurry Format) - Protocol (50 prep) APPLICATION NOTES Isolation of DNA From As Little As 25 uL of Urine Using Norgens Urine DNA Isolation Kit Isolation of High Quality Urinary DNA Using Norgen’s Urine DNA Isolation Kit Determination of the DNA Molecular Weight (MW) from different Norgen Columns and Isolation Methods SAFETY DATA SHEETS (18100) Urine DNA Isolation Kit - Safety Data Sheet (48800) Urine DNA Isolation Kit (Slurry Format) - Safety Data Sheet (50100) Urine DNA Isolation Maxi Kit (Slurry Format) - Safety Data Sheet POSTERS Genomic Profiling of Hepatitis B Virus and Hepatitis C Virus From 1 mL of Urine - Asm 2007 A Novel Method To Capture Methylated Human DNA From Urine Implications For Prostate Cancer Screening - Ascb 2008 The Urinary Genomic and Proteomic Profiling of Hepatocellular Carcinoma Patients Infected With Hepatitis C Virus - ASM 2008 Sensitivity of DNA Extraction Methods from Different Bodily Fluids for Human Identification Citations Powered by Bioz See more details on Bioz
Urine DNA Isolation Kits Fast and reliable purification of genomic and apoptotic DNA from urine For research use only and NOT intended for in vitro diagnostics. CE-IVDD marked diagnostic slurry format available here Powered by Bioz Urine DNA Isolation Kits Fast and reliable purification of genomic and apoptotic DNA from urine SKU 18100 Format Spin Column Slurry Maxi Slurry Size 50 Preps Contact Distributor