DNA extraction from human urinary sediment

Abstract

DNA was extracted from urinary sediments and was sufficient for polymerase chain reaction (PCR) and enzymatic analysis, even if DNA from microorganisms coexisted. From urine samples, the yield of DNA ranged from trace levels to 20 micrograms per 10 mL urine. When urinary sediment was stored in ethanol, DNA remained stable for 2 weeks or more. Individual identification and sex determination could easily be performed using either fresh or ethanol-fixed urine. In conclusion, urine can be used as a source for PCR-based investigations and genetic studies.

Urine — a source for noninvasive genetic monitoring in wildlife

“…However, species-and loci-related differences need to be considered in such a comparison. In addition, urine contains predominantly DNA of the correspondent individual (Yokota et al 1998), and therefore, no dietary remains affect genetic analyses (see Murphy et al 2003).…”

PCR-ready human DNA extraction from urine samples using magnetic nanoparticles

“…Further, the methodology described requires no hazardous reagents (such as phenol and chloroform) during the entire process. Finally, the entire CMNP extraction process required less than 30 min, a significant time savings over the traditional phenol/chloroform method, which needed at least several hours to complete [9,10].…”

“…In a control study, gDNA was isolated from pelleted urine cells (1500 × g, 5 min) using a modified phenol/chloroform method [10]: pelleted cells were washed with TNE buffer (10 mM Tris, 100 mM EDTA and 100 mM NaCl, pH 8.0) and re-centrifuged followed by the addition of 600 l of lysing solution (1% SDS in TNE buffer). The mixture was incubated for 1 h at 55 • C, followed by the addition of proteinase K to a final concentration of 100 g/ml.…”

“…The isolation of DNA from urine, rather than traditional blood samples is advantageous, since it reduces the pain and risk of infection associated with venous blood collection, particularly during athletic competitions, where urine is the least invasive, and potentially the only source of DNA available for identifying individuals. The small amounts of nucleated cells (epithelial, leukocytes, and even exfoliated malignant cells) and cell-free DNA present in voided urine makes diagnosis (e.g., K-ras for cancer), genotyping and screening possible [5][6][7][8][9][10].…”

“…Current gDNA extraction methods use centrifugation or filtration, which are very time-consuming and labor-intensive processes that cannot be automated. Further, existing approaches require large volumes of urine to obtain sufficient cells, and employ toxic organic reagents, enzymes, and spin columns to isolate DNA from proteins and other contaminants [1,[8][9][10][11].…”