Principle:
Precautions:
The QIAamp DNA Blood Mini Kit combines the selective binding properties of a silica-gel-based membrane with the speed of a microspin. Purification requires no phenol/chloroform extraction or alcohol precipitation, and involves very little handling. DNA is eluted in Buffer AE or water, ready for direct addition to PCR or other enzymatic reactions.
- Mix buffer AL thoroughly by shaking before use. If a precipitate has formed dissolve by incubating at 56°C.
- Avoid touching the QIAamp membrane with the pipet tip.
- Wear powder-free gloves throughout the entire procedure.
- Pipet 20 ul QIAGEN Protease into the bottom of a 1.5 ml microcentrifuge tube.
- Add 200 ul sample into the tube.
- Add 200 ul Buffer AL (with 6 ul of Internal control) to the sample. Mix by pulse-vortexing for 15 seconds.
- Incubate at 56 deg C for 10 min.
- Briefly centrifuge the tube to remove drops from the inside lid.
- Add 200 ul ethanol to the sample and mix again by pulse-vortexing for 15 seconds. Briefly centrifuge the tube to remove drops from the inside lid.
- Carefully apply mixture from step 6 to the QIAamp spin column, close the cap and centrifuge at 8,000 rpm for 1 min. Place the spin column in a clean 2 ml collection tube and discard the tube containing the filtrate.
- Carefully open the column and add 500 ul Buffer AW1 and centrifuge at 8000 rpm for 1 min. Place the spin column in a clean 2 ml collection tube and discard the tube containing the filtrate.
- Carefully open the spin column and add 500 ul Buffer AW2. Centrifuge at 14,000 rpm for 3 min.
- (Optional) Place the spin column in a new collection tube and centrifuge at 14,000 rpm for 1 min.
- Place the spin column in a new 1.5 ml microcentrifuge tube and discard the collection tube containing the filtrate. Open the spin column and add 60 ul Buffer AE, incubate at room temperature for 5 min and then centrifuge at 8,000 rpm for 1 min. The filtrate now contains the DNA, ready for PCR.
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