PCR-ready-Reagent (DNA-Purification from blood-samples)
The PCR-ready DNA Purification System provides a simple, effective, safe and inexpensive approach to isolate DNA from whole blood for amplification analysis. The technique is based on the thermocoagulation of proteins, polysaccharides and some other biomolecules under specific conditions
simple - effective - inexpensive
| Catalog no. | Pack size | Price, Euro | Shop |
| 805060 | 50 preps | 109,00 | Add to cart |
| 805120 | 100 preps | 199,00 | Add to cart |
Protocol:
1. Collect 2 ml of blood stabilized with EDTA or citrate. Avoid use of Heparin as
anticoagulant!
2. Use the blood during 2 hours after collection without cooling. Do not store store
the blood-sample at 4 – 8 °C longer than 24 hours for the DNA-Purification
procedure.
3. Before DNA purification mix the blood if visible layers appeared. Transfer 1 ml of
the blood into the 1,5 ml Test-tube with locked cap.
4. Close the tube and centrifuge at 3000 rpm for 5 min at room temperature. The
blood will be separated to 3 visible layer (red cells below the plasma above and
intermediate film of leucocytes).
5. Carefully remove plasma by pipette with no damage to leukocyte film. Do not try
to remove plasma completely otherwise the removal of leukocytes will be
inevitable.
6. Close the tube and incubate it for 1 hour at –20ºС (in a freezer) .
7. Defreeze the content of the tube completely at room temperature.
8. Measure (roughly) the volume of the tube content (as usual it is 550 microliters).
Add equal volume of PCR-ready-Reagent into the tube. Close the tube with the
lock.
9. Vortex well for 10 seconds.
10. Place the tube in the preliminary heated-up thermostat at 98ºC, incubate for 15
min.
11. Place the tube into the centrifuge with the lock oriented to the axis of the rotor.
Centrifuge the tube 8000-14000 rpm 20-30 sec at room temperature.
12. Use the supernatant for PCR reaction. As a rule 5 microliters of the supernatant is
enough for the successful PCR.
The supernatant can be transferred to the new tube and stored up to 6 months at
-20°C.
Important Notes.
1. Do not use Blood samples which are stabilized with Heparin as anticoagulant!
2. Do not use coagulated Blood for DNA Purification.
3. Try to avoid damage of leucocyte film in Step 5. This step is very important since
removal plasma together with leucocytes will result in completely unsuccessful
procedure.
4. For themocoagulation in step10 always use Test-tubes with Safe-Lock!
