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Quick-DNA™ Urine Kit

Purify cellular and/or cell-free DNA easily and reliably from up to 40 ml of urine.
Uniquely formulated urine conditioning reagent allows stabilization of DNA in urine for up to 1 month at ambient temperature.
Zymo-Spin™ column technology ensures DNA is ready for all sensitive downstream applications including qPCR, DNA sequencing, arrays, and DNA methylation analysis.

Product Size Catalog # Price Qty
Quick-DNA™ Urine Kit 50 Preps D3061
$417.00

About Quick-DNA™ Urine Kit

The Quick-DNA™ Urine Kit is an innovative product designed for easy, reliable, and rapid isolation of cellular and/or cell-free DNA from up to 40 ml of urine. The product features a uniquely formulated urine DNA stabilization reagent that also functions as a precipitation reagent. After collection, total (cellular and cell-free) or cell-free urine can be stored at ambient temperature for up to one month by adding the Urine Conditioning Buffer. When ready to extract the urine DNA, just add the Clearing Beads, vortex, and centrifuge to collect the precipitate. Following precipitation, chemical lysis and enzymatic digestion are used to extract DNA from the precipitate. The DNA is subsequently purified and concentrated using Zymo-Spin™ IC-S Columns. Urine DNA isolated with the Quick-DNA™ Urine Kit is ideal for qPCR, array, methylation analysis, and other downstream applications.


DNA Quality High quality DNA can be used for downstream application, such as PCR, bisulfite treatment/methylation detection, array, etc.
Sample Sources Urine.
DNA Yield The DNA binding capacity of the column is 5 μg. Note that DNA yield may vary depending on the urine itself. Female urine typically yields more DNA than male urine. Urine DNA from healthy female individual ranges on average from 6 - 1000 ng/ml. DNA from healthy male individual ranges on average from 2 - 20 ng/ml.
Equipment Needed A centrifuge, a microcentrifuge, and a heat block / water bath.
Recommended Sample Volume Up to 40 ml of urine per standard preparation. The sample volume can be scaled up or down if required.
DNA Size Capable of recovering DNA fragments from 100 bp to 23 kb.

   

DNA yields increase linearly with increasing volumes of urine from healthy subjects extracted using the Quick-DNA™ Urine Kit. DNA was isolated from 1 ml, 10 ml, 25 ml, and 40 ml urine. DNA concentration was quantified using the Femto™ Human DNA Quantification Kit (Zymo Research, E2005).

Both cellular and cell-free DNA can be effectively purified from urine using the Quick-DNA™ Urine Kit. 5 ml of urine from a healthy female donor was processed and DNA was eluted in 20 μl final volume. 1 μl of the sample was analyzed using a 2200 Tapestation.

Step 1

(A) Total (cellular and cell-free) DNA Precipitation

Transfer up to 40 ml urine2 into a microcentrifuge tube or a conical tube.

Note: Use a microcentrifuge tube if processing ≤ 1 ml urine.
 Use a 15 ml conical tube if processing > 1 ml to14 ml urine.
 Use a 50 ml conical tube if processing > 14 ml to 40 ml urine.

Step 2

Add 70 μl Urine Conditioning Buffer for every 1 ml of urine (e.g. Add 350 μl Urine Conditioning Buffer to 5 ml urine). Mix the urine mixture well by vortexing.

After adding and mixing urine with Urine Conditioning Buffer, urine can be stored up to 1 month at ambient temperature. At time of processing, mix the urine mixture well by vortexing and continue to the next step.

Step 3

Add 10 μl Clearing Beads3 if processing ≤ 14 ml urine and add 20 μl Clearing Beads3 if processing >14 to 40 ml urine. Mix the urine mixture well by vortexing.

Step 4

Centrifuge at 3,000 x g for 15 minutes.
(If isolating DNA from gram (+) bacteria, fungal, yeast, or other tough-to-lyse samples in urine, continue to Appendix B on page 7)

Step 5

(B) Protein Digestion

Without disturbing the pellet, slowly decant or pipette out the urine supernatant leaving behind 100 - 400 μl of pellet.

Recommended: leave at least 200 μl of pellet if processing 15 - 40 ml urine.

Note: you could adjust the volume to 100 - 400 μl by adding DNase/RNase-Free Water to the pellet.

Step 6

Add an equal volume of Urine Pellet Digestion Buffer to the pellet (e.g. Add 100 μl Urine Pellet Digestion Buffer to 100 μl pellet). Resuspend the pellet well by pipetting or vortexing.

Step 7

Add 5% (v/v) of Proteinase K to the resuspended pellet (e.g. Add 10 μl Proteinase K to 200 μl mixture), mix well by vortexing, and incubate the pellet mixture at 55 ˚C for 30 minutes.

Step 8

(C) DNA Purification

Add 1 volume of Genomic Lysis Buffer to the digestion mixture (e.g. Add 210 μl Genomic Lysis Buffer to 210 μl digestion mixture) and mix well by vortexing.

Step 9

Transfer the sample into a Zymo-Spin™ IC-S Column in a Collection Tube. Centrifuge for 1 minute. Discard flow-through. Repeat with the remaining mixture1.

Note: The column may need to be loaded multiple times. Transfer 800 μl maximum for each load of mixture.

Step 10

Transfer the Zymo-Spin™ IC-S Column to a new Collection Tube.

Step 11

Add 200 μl of Urine DNA Prep Buffer to the spin column. Centrifuge for 1 minute. Discard flow-through.

Step 12

Add 700 μl Urine DNA Wash Buffer to the column and centrifuge for 1 minute. Discard flow-through. Repeat this step with 200 μl Urine DNA Wash Buffer.

Step 13

Transfer the spin column to a DNase/RNase-free microcentrifuge tube. Add ≥ 10 μl2 DNA Elution Buffer3 or water directly on the column matrix and let stand for 3 - 5 minutes at room temperature. Centrifuge at full speed for 1 minute. The eluted DNA can be used immediately or stored at ≤ -20 ˚C for future use.

Note: If the concentration of the DNA is > 100 ng/μl in the first elution. Most of the DNA will be recovered on the first elution however the loading eluate from the first elution back onto the column, incubating for 3 - 5 minutes at room temperature, and centrifuging again can increase total yield

For specific notes and additional information, please see the product protocol PDF.
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Featured Citations

DNA was extracted from urinary epithelial cells, non-invasively available tissue, using the Quick-DNATM Urine Kit in study of the Old Order Amish population in the US suspected of having mitochondrial DNA mutations. Collected urine samples were assessed for heteroplasmy level for the m.3243A>G mutation using high-resolution melt (HRM) profiling and confirmed through DNA Sanger sequencing. This finding represents the first report of mitochondrial respiratory chain disorders in the Amish community and suggests that it may be under-diagnosed.
Researches used the Quick-DNA™ Urine Kit (previously known as Extract-all Urine DNA kit) to detect and investigate the role of tumor cell-free DNA copy number variations (CNVs) in urine as clinically relevant diagnostic and predictive biomarkers for metastatic hormone sensitive and metastatic castrate resistant stages of prostate cancer. Urine cfDNAs derived from Quick-DNA™ Urine Kit were used to perform whole genome sequencing to generate high quality sequencing library. Researchers were able to detect CNVs in all samples, furthermore they were able to detect specific urine cfDNAs genomic aberrations at loci of PTEN, TMPRSS2, AR, and NOTCH1. CNV changes after treatment (androgen deprivation therapy and doxetacel chemotherapy) allowed for the identification of treatment-associated CNV changes, most important of which have been reported to be aberrant in prostate cancer such as copy number changes in RNF43 and ZNRF3 loci and in the LGR4 and MYC proto oncogene loci. These findings could prove potential predictive and prognostic classifiers for advance prostate cancer patients.

The Zymo Kit and more specifically the Cell-DNA method presents as the best option to improve the outcome of our LR-PCR method.
R. D. (University of Sydney and Royal North Shore Hospital)
We tested it this week and it worked beautifully. We really would like to order this kit to process more samples!
R. D. (Medical College of Wisconsin)
The protocol was easy to use/follow and it didn't take a great deal of time to get results.
S. H. (South College School of Pharmacy)
Urine DNA isolation kit is very much needed and I like Zymo kits for the simplicity an reliability.
A. C. (Sanford-Burnham Medical Research Institute)
The kit is very useful as it lets us to analyze big volumes without an ultracentrifugation step.
S. B.M. (University of Barcelona)
Good yields of DNA in a very simple efficient process.
R. W. (Medical University of South Carolina)
Click here to submit your review of the Quick-DNA™ Urine Kit.