Urine DNA Isolation Micro Kit

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 lane 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.