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title: "TheNessLab : RNA Extraction with Maxwell RSC Plant RNA Kit (Promega)"

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TheNessLab : RNA Extraction with Maxwell RSC Plant RNA Kit (Promega)

Created by Eniolaye Balogun on Nov 25, 2020

Prep Homigenization Solution (Note: This mixed solution ONLY lasts for 30 days, so ensure you make just enough) 

• Add 20 ul of refrigerated 1-Thioglycerol to 1 mL of Homogenization Solution

Prep DNase I

• Add 275 uL of Nuclease-free water to the entire vial of lyophilized DNase I (Mix well) 
• Add 5 uL of Blue Dye to mixture 

Procedure

1. Prep your homogenization solution and ensure it has been placed on ice for at least 30 minutes
2. Store the TissueLyser pods at -80*C fridge for minimum 30 minutes 
3. Label 2 mL Erlenmeyer tubes per sample and place in TissueLyser pods 
4. Add 200 ul of homogenization solution into each sample
5. Shake your mixture at 25 shakes/s for 30 seconds 
6. Quickly place samples on ice 
7. Add 200 ul of Lysis buffer per tube on ice 
8. Immediately vortex at high for 15-30 sec (until the pellet is fully broken) 
9. Incubate all samples at room temperature for 15 minutes 
10. Centrifuge at 16 K CFU for 2 mins 
11. Pour supernatant in well#1
12. Add 5 uL of prepared DNase I to well #4 (It should turn green over time if done correctly)
13. Add 50 uL of nuclease-free water to the elution tube 
14. Add plunger and run on Maxwell RSC Instrument 
15. If your run is aborted, follow instructions by the machine

Document generated by Confluence on May 22, 2024 11:44

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