GALENVS magneti Plant RNA Extraction Kit User Guide

  GALENVS Magneti Plant RNA Extraction Kit

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. Avoid pellet, transfer up to 300–400µl of supernatant to clean the centrifuge tube. Add 1000µl of Binding Buffer, vortex for 10–20s, and wait 5 mins.
4. Place the tube on a magnetic rack to capture. Wait 1 min then discard the supernatant.
5. Add 600µl of Wash Buffer #1 to the tube and vortex for 10–20s. Wait 1 min then place the tube on a magnetic rack to capture. Wait 1 min then discard the supernatant.
6. Add 600µl of Wash Buffer #2 to the tube and vortex for 10–20s. Place the tube on a magnetic rack to capture. Wait 1 min then discard the supernatant.
7. Add 100µl of Elution Buffer to the tube and mix briefly. Wait 1 min. Place the tube on a magnetic rack to capture.
8. Wait 1 min then transfer the supernatant to the clean microfuge tube.
9. Add 100µl of DNase Reaction Buffer to the DNase pellet and then add 100µl of glycerol. Mix by gently inverting the tube. The reconstituted pellet must be stored at -20 °C.
10. For each RNA sample, prepare DNase buffer by adding 10µl of DNase prepared in the previous step to 40µl of DNase Reaction Buffer in a microfuge tube.
11. Add 50µl of DNase buffer to 50µl of RNA sample in a microfuge tube. Gently shake the mix for the 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 the microfuge tube and vortex for 10s. Incubate the mixture for 5 mins at room temperature.
14. Place the tube on a magnetic rack for 2 mins. Discard supernatant. Mix DNase with buffer by gently inverting the tube a few times.
15. Resuspend beads in 600µl Wash Buffer #3. Vortex for 10–20s. Place the tube on the magnetic rack, wait for 1 min, and discard the supernatant.
16. Repeat the washing step with Wash Buffer #3.
17. Dry beads on a magnetic rack for 2 mins after the third wash. Remove any wash buffer left at the bottom of the tube at the end of the drying step.
18. Add 50µl of the Elution Buffer to the tube and mix briefly.
19. Place the tube on the magnetic rack, wait for 1 min then transfer the supernatant to the clean tube.

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