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DNA and RNA extraction Kit Protocol
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The procedure instructions for the extraction kit are very simple; allowing its users to purify DNA or RNA from a wide variety of target sources in as little as 30 minutes. In fact, it is as follows:
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1. Transfer 150 (300) μl of plasma, serum, urine, cell culture supernatant, cell-free fluid or virus infection into tissue or cell in the 1.5 ml microcentrifuge tube.
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If the volume of the sample is less than 150 L, the sample should be set to 150 L with WATER treated with DEPC. If the volume of the sample is higher, 15 ml. centrifugal tube should be used.
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2. Add 300 (600) μl lysis buffer. If the volume changes, vary the quantity.
3. Mix by stirring in vortex for 15 seconds.
4. Incubate at room temperature (15-25 °C) for 10 minutes.
5. Add 300 (600) μl joining buffer and mix gently.
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If the sample volume is greater than 150 He, we should increase the amount of bonding buffer.
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6. Place a rotating column in a 2 ml collection tube provided.
7. Load the lyses into the column and centrifuge at 13,000 rpm for 1 minute.
8. Dispose of the solution in the collection tube and place the column back in the same 2 ml. collection tube.
9. Add 500 μl of wash buffer A, to the column and centrifuge for 1 minute at 13,000 rpm.
10. Dispose of the solution in the collection tube and place the spin column back into the same 2 ml. collection tube.
11. Add 500 μl of wash buffer B to the column and centrifuge for 1 minute at 13,000 rpm.
12. Discard the solution in the collection tube and place the spin column back into the same 2 ml. Centrifuge collection tube for 1 minute at 13,000 rpm.It is important to dry the membrane as residual ethanol may interfere with downstream reactions.
13. Place the column in a 1.5 ml microcentrifuge tube without RNasa (not included), and add 30 – 60 l of elution buffer directly onto the centrifuge column membrane. It is important to dry the membrane as residual ethanol may interfere with downstream reactions. Do not touch the membrane with the tip of the pipette.
14. Incubate at room temperature for 1 minute, then centrifuge for 1 minute at 13,000 rpm.
15. Use 2-5 μl of elued solution as a template for PCR or RT-PCR.
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In case you are interested or interested in purchasing this product, please contact us, Durviz.