Catalog

18100

Urine DNA Isolation Kit

( 50 preps )
Norgen Biotek

588.66$

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Additional description

Urine DNA Isolation Micro Kit (Cat# 18100 )

Fast and reliable purification of genomic and apoptotic DNA from urine

  • Rapid isolation of small and large species of DNA from urine
  • Convenient spin column format
  • Purified DNA is highly suited to sensitive downstream applications

Product Description

Norgen’s Urine DNA Isolation Micro Kit provides afast, reliable and simple procedure for isolating DNA from as low as 50 µL of urine up to 2 mL of urine. Many advantages favour the use of urinary DNA for cancer biomarker discovery and diagnosis of different pathogens over the use of blood and tissue samples. 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. Both types of DNA can be isolated reliably using this kit.Typical yields of DNA isolated will vary depending on the input sample, with more concentrated samples tending to yield more DNA.

Purification is based on spin column chromatography using Norgen's proprietary resin as the separation matrix. Briefly, urineDNA isbound to the column in the presence of Binding Solution (BIND).This is folllowed by washing of the bound DNA to remove the remaining proteins or other impurities (WASH).Lastly, the purified total urine DNAis eluted into the provided Elution Buffer (ELUTE). Please see the procedure flowchart to the right.

The kit allows for the removal of highly concentrated salts, metabolic wastes and proteins, to allow for the isolation of high quality, concentrated DNA. Preparation time for a single sample is about 30 minutes. The purified urine DNA is compatible with most of the molecular biological applications such as PCR, q-PCR and DNA fingerprinting

FEATURES AND BENEFITS

  • Fast processing - Rapid spin-column format allows for the processing of multiple samples in 30 minutes.
  • Isolation of both types of urine DNA - Isolate 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).
  • High quality DNA - Removal of highly concentrated salts, metabolic wastes and proteins provides high quality, concentrated DNA to be used in various downstream applications.
  • Recovered DNA is suitable for downstream applications - Purified DNA is fully compatible with PCR, q-PCR and DNA fingerprinting

Specifications

The purified urine DNA is compatible with most of the molecular biological applications such as:

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 DNAwas isolated from two different 1.5 mL urine samples.The samples were processed according to the bind, wash andelute procedure provided in the kit, and theDNA waseluted into2 separate elutionvolumes. The proteins were first eluted into 100 µL of Elution Buffer (E1), followed by a second elution using75 µL of Elution Buffer (E2).The purified urine DNAwas then run on a1.2% agarosegel, andeachlane 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 is10 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 mLurine 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 usedas the Template inPCR Reactions. Total urinary DNA was isolated from three different 1.5 mLurine 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.

APPLICATIONS

Kit Specifications
Volume of Urine Processed
2 mL
Size of DNA Purified
Large (>1kb) and small (150-
250 bp)
Average Yield
Up to 50 ng
Time to Complete 10 Purifications
40 minutes
* Yield will vary depending on the type of sample processed

Urine DNA Isolation Kit Contents
1. Activation Buffer
2. Binding Solution I
3. Binding Solution II
4. Lysis Solution
5. Pronase
6. Proteinase K
7. Elution Buffer
8. Wash Solution
9. Micro Spin Columns
10.Collection Tubes
11.Elution Tubes
12.Product Insert


Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 1 year in their unopened containers. The Pronase and Proteinase K should be stored in aliquots at -20°C upon reconstitution. These products are stable at room temperature in their lyophilized form.