Description
The One Tube DNA Purification system provides a simple, fast and effective approach to isolate genomic DNA of a wide range of bio-samples as epithelium from urethra, cervix, vagina, buccal epithelium, sperm, prostate liquid, urine precipitates, saliva, phlegm, synovial fluid etc. For qPCR and Real Time applications the OTP -Reagent is also available without stain.
The technique based on the thermocoagulation of proteins, polysaccharides and some other biomolecules under specific conditions. The process takes less than half an hour. The resultant DNA can be effectively used in PCR .DNA Purification system is a clever transportation-storage media for bio-samples.
Storage conditions
Protocol for sample preparation
Important: The amount of cells (if visible) should not be more than approx. 3 mm in diameter.
A. Transfer epithelial cells from urethra, cervix, vagina, buccal epithelium with a disposable sterile swab into the tube with OTP -Reagent. Remove the swab from the tube, close the tube. Remove mucus excess from cervix before sampling.B. Urine pellet Urine should be collected early in the morning. transfer to the laboratory in 1-3 hours after collection. Urine cannot be cooled down or frozen, it cannot be stored longer than several hours. Mix urine, take out 10-15 ml into the centrifuge tube, centrifuge 10-15 min at 3000 rpm at room temperature. Discard supernatant carefully, not disturbing the pellet transfer 0.5 ml of the pellet suspension into 1.5 ml tube centrifuge at 12000 rpm for 15 sec at room temperature. Remove supernatant carefully. (Option: wash the pellet once with 1 ml of saline solution) Add total volume of OTP -Reagent to the pellet in the 1.5 ml tube, mix well by pipetting and transfer the suspension back to original OTP -Reagent tube C. Sperm, prostate fluid, saliva, phlegm, synovial fluid. Transfer 20 µl of the sample into the tube with OTP -reagent.
DNA purification for PCR analysis Close and check OTP -tubes with the bio-sample carefully Vortex the tubes with the mixture well Place the tubes into pre-heated (98°C) dry thermostat Incubate for 15 min. Let cool down at room temperature 1-2 min Centrifuge tubes at 12000 rpm for 15 sec Use the supernatant as a template for the subsequent PCR , depending on your protocol. Note: If PCR is inhibited by some extra impurities in the sample, transfer 100 µl of supernatant into the new tube with 200 µl of OTP -Reagent and repeat the steps 1-3.
Important Notes
Do not use transport media different from OTP -Reagent for the samples – the components of the transport media may cause inhibition of PCR reaction! Try to avoid displacement of mucus, blood, pus and other contaminating material into the tube with OTP -Reagent Try to avoid anesthesia if possible. Some medicines may cause inhibition of PCR . Do not overload the tube with the excess of epithelium cells! The amount of epithelial cells (if visible) should not be more than approx. 3 mm in diameter. In some cases, when the excess of material is obvious, add additional 200 microliters of OTP -Reagent to the sample.
