9
16. Leave the tube on the magnetic separation device for 3 minutes to air dry the Mag-
Bind® Particles CNR. Remove any residual liquid with a pipettor.
17. Prepare the Mag-Bind® DNase I digestion mix as detailed in the table below:
Note:
If total nucleic acid (DNA and RNA) is desired, skip Mag-Bind® DNase I
digestion steps (Steps 18-24) and proceed to Step 25 for isolating both DNA and
RNA.
Number of
Samples
Mag-Bind® DNase I
Digestion Buffer
Mag-Bind® DNase I
Total Volume
1
49 µL
1.0 µL
50 µL
4
215 µL*
4.4 µL*
219.4 µL
10
539 µL*
11 µL*
550 µL
96
5.18 mL*
106 μL*
5.29 mL
* Volumes are calculated 10% extra to offset pipetting error.
Important Notes:
•
Mag-Bind® DNase I is very sensitive and prone to physical denaturation.
Do not
vortex the Mag-Bind® DNase I mixture.
Mix gently by shaking the plate.
•
Freshly prepare Mag-Bind® DNase I digestion mix right before RNA isolation.
•
All steps must be carried out at room temperature. Work quickly, but carefully.
18. Add 50 µL Mag-Bind® DNase I digestion mix. Pipet up and down 20 times or shake
gently for 2 minutes to mix.
Note:
It is very important to remove any liquid drop from the wells before adding
the Mag-Bind® DNase I digestion mix. Mag-Bind® DNase I digestion mix must be used
immediately once it is prepared.
19. Let sit at room temperature for 10-15 minutes.
20. Add 200 µL RNA Wash Buffer II. Pipet up and down 20 times or shake for 5 minutes to
mix.
M2837 Mag-Bind® Blood RNA 96 Protocol