About ZR Urine DNA Isolation Kit™
The ZR Urine DNA Isolation Kit™ is an innovative product designed for the easy, reliable, and rapid isolation of total DNA from cells and biological sediment in urine samples. The product enables isolation of cells from urine using a syringe fitted with a uniquely-designed syringe filter. Following separation, cells are lysed and the collected lysate can be processed immediately or at a later time following transportation and/or storage. The DNA isolation procedure is simple and can be performed in less than 10 minutes with the technologies featured in this kit. Total DNA isolated with the ZR Urine DNA Isolation Kit™ is ideal for PCR, array, methylation detection, etc.
|Equipment||Syringe (30 ml, 1 ml), Microcentrifuge|
|DNA Size Limits||100 bp to ≥ 40 kb|
|DNA Purity||High quality DNA for PCR, bisulfite treatment/methylation detection, array, etc., is eluted with DNA Elution Buffer or water. (A260/A280>1.8)|
|Sample Size||30 ml per standard preparation. The sampled volume can be scaled up or down if required.|
|DNA Yield||The DNA binding capacity of the column is 5 µg. Female urine typically yield 14-200 ng DNA/ml. Male urine yield 4-60 ng DNA/ml.|
DNA purified from human urine using the ZR Urine DNA Isolation Kit™ is ideal for use in PCR. The gel image above shows the results of PCR amplification of 15 ng, 1.5 ng, and 0.15 ng total DNA (1x, 10x 100x dilutions, respectively) isolated from human urine using primers specific for the human ß-actin gene. (M) is a 50 bp DNA ladder (Zymo Research) and amplicons are indicated (?). The (+) and (-) are positive and negative controls, respectively.
Take up fresh urine into a 30 ml syringe. Attach a provided ZRC GF™ Filter to the syringe and push the urine completely through the filter into an appropriate waste container2. Remove residual urine from the filter by pushing through several volumes of air.
Using a 1 ml syringe, push 1 ml of Genomic Lysis Buffer through the filter and collect the flow-through into a Zymo-Spin™ IC Column in a Collection Tube3. Cap the column and invert assembly gently several times. Centrifuge at ≥ 10,000 x g for 1 minute. Discard the flow-through from the Collection Tube.
Add 200 µl of DNA Pre-Wash Buffer to the spin column. Centrifuge at ≥ 10,000 x g for 1 minute. Discard flow-through.
Add 500 µl g-DNA Wash Buffer to the column and centrifuge at ≥ 10,000 x g for 1 minute.
Transfer the spin column to a 1.5 ml microcentrifuge tube. Add ≥ 6 µl DNA Elution Buffer or water directly to the column matrix. Wait for 1 minute then centrifuge at ≥ 10,000 x g to elute the DNA.
The eluted DNA can be used immediately or stored at ≤ -20°C for future use.