GALENVS PR0050-QSG Plant RNA Extraction Kit User Guide

GALENVS PR0050-QSG Plant RNA Extraction Kit

Instruction For Use

1. Add up to 50mg of ground fresh plant leaves to the lysis bead tube provided.

2. Add 600µl of Lysis Buffer and 60µl PPB. Mix for 3 mins using TissueLyser at max speed or vortex for 10 mins; then centrifuge at 20,000g for 2 mins.

3. Avoiding pellet, transfer up to 300–400µl of supernatant to clean centrifuge tube. Add 1000µl of Binding Buffer #1, vortex for 10–20s, and wait 5 mins.

4. Place tube on magnetic rack to capture. Wait 1 min then discard supernatant.

5. Add 600µl of Wash Buffer #1 to the tube and vortex for 10–20s.
   Wait 1 min then place tube on magnetic rack to capture. Wait 1 min then discard supernatant.
   Repeat this step

6. Add 600µl of Wash Buffer #2 to the tube and vortex for 10–20s.
   Place tube on magnetic rack to capture. Wait 1 min then discard supernatant.
   Repeat this step.

7. Add 100µl of Elution Buffer to tube and mix briefly. Wait 1 min.
   Place tube on magnetic rack to capture.

8. Wait 1 min then transfer supernatant to clean microfuge tube..

9. Add 2.5ml of DNase Reaction Buffer to DNase pellet.
   Mix by gently inverting the tube.

10. Add 50µl of DNase buffer to 50µl of RNA sample in a microfuge tube.

11. Gently shake the mix for 20s, and incubate at room temperature for 25 mins.

12. Mix 40µl of Binding Beads with 400µl Binding Buffer #2.

13. Add 400µl of Binding Bead and Binding Buffer #2 mixture to microfuge tube and vortex for 10s.
    Incubate the mixture for 5 mins at room temperature.

14. Place tube on magnetic rack for 2 mins. Discard supernatant.

15. Resuspend beads in 600µl Wash Buffer #3. Vortex for 10–20s.
    Place tube on magnetic rack, wait for 1 min and discard supernatant.

16. Repeat washing step with Wash Buffer #3.

17. Dry beads on magnetic rack for 2 mins after the third wash.
    Remove any wash buffer left at the bottom of tube at the end of drying step.

18. Add 50µl of Elution Buffer to tube and mix briefly

19. Place tube on magnetic rack, wait for 1 min then transfer supernatant to clean tube.

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