One-Tube DNA purification (OTP-Reagent)
| Catalog no. | Pack size | Price, Euro | Shop |
| 805050 | 50 preps | 109,00 | Add to cart |
| 805100 | 100 preps | 199,00 | Add to cart |
| 806050 (for qPCR) | 50 pres | 119,00 | Add to cart |
| 806150 (for qPCR) | 100 preps | 199,00 | Add to cart |
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 and can be stored at +2-8°C for one week or up to 6 months at -20°C.
The One-Tube DNA Purification System is a clever transportation- storage media for bio-samples.
Storage Conditions: The OTP-Reagent should be stored at +2-8°C for 1 year or can be stored and transported at room temperature (up to 25°C) not longer than 5 days.
Protocol for sample preparation
1. Collection of biosamples.
Important: The amount of cells (if visible) should be not more than approx. 3 mm in diameter.
A. Transfer Epithelial cells from urethra, cervix, vagina, buccal epithelium with the 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.
1. Urine should be collected early in
the morning.
2. transferred to the laboratory in
1-3 hours after collection.
3. Urine cannot be cooled down or frozen, it cannot be stored longer than several
hours.
4. Mix urine, take out 10-15 ml into the centrifuge tube, centrifuge 10-15 min
at 3000 rpm at room temperature.
5. Discard supernatant carefully, not disturbing the pellet
6. transfer 0,5 ml of the pellet suspension into 1,5 ml tube
7. centrifuge at 12000 rpm for 15 sec at room temperature.
8. Remove supernatant carefully.
9. (Option: wash the pellet once with 1 ml of saline solution)
10. 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. 20 microliters of the sample
transfer into the tube with OTP-reagent.
DNA purification for PCR analysis
1. Close OTP-tubes with the bio-sample carefully and check
2. Vortex well the tubes with the mixture
3. Place the tubes into the preliminary heated up to +98oC dry thermostat
4. incubate for 15 min.
5. Let cool down at room temperature 1-2 min
6. Centrifuge tubes at 12000 rpm for 15 sec
7. 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.
The supernatant can be stored at +2-8°C for one week or up to 6 months at -20°C.
Important Notes.
1. Do not use transport media different from OTP-Reagent for the samples – the
components of the transport media may cause inhibition of PCR reaction!
2. Try to avoid displacement of mucus, blood, pus and other contaminating material into the
tube with OTP-Reagent
3. Try to avoid Anesthesia if possible. Some medicines may cause inhibition of PCR.
4. Do not overload the tube with the excess of epithelium cells! The amount of epithelial
cells (if visible) should be not 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.
