RNA extraction is a process to take out the Viral RNA. RNA Extraction is one of the most important parts of Coronavirus Testing. Before going to Extract the RNA you have to Decontaminate the Simple with Lysis Buffer. As we know Different Kit have different Procedure but the mechanism is the same for all. Today we are going to discuss the procedure of QIAGEN Viral RNA Kit Step by Step
RNA Extraction Procedure for Corona Virus-Step by Step Procedure
- Pipet 560 µl of prepared Lysis Buffer containing carrier RNA in a 1.5 ml microcentrifuge tube
- Add 140 µl of Sample to the tube containing Lysis Buffer-Carrier RNA Mix properly by vortexing (This Step should be Done inside the Biosafety Cabinet (Class II) with proper PPE)
- Incubate at room temperature for at least 10 minutes (More than 10 min incubation will not any effect on Viral RNA)
- After incubation quick spin the tube in a centrifuge to removes drops from the inside of the lid
- Add 560 µl ethanol (96-100%) to the sample and mix properly by vortexing. After mixing briefly centrifuge the tube
- Transfer 630 µl of the solution from 1.5 ml microcentrifuge tube to the 2 ml mini Column (Collection Tube). Close the Cap and Centrifuge at 8000 rpm for 1 min then place the mini-column into fresh 2 ml collection tube and discard the filtrate solution tube
- Repeat the same step for remaining solution
- Add 500 µl of AW1 wash buffer and centrifuge at 8000 rpm for 1 min. Place the mini-column into the fresh collection tube
- After wash buffer 1 carefully open the cap and add 500 µl of AW2 wash buffer and centrifuge at 14000 rpm for 3 min. Place the mini-column into the fresh collection tube
- Dry spin at centrifuge at 8000 rpm for 1 min.
- Place the mini-column into fresh 1.5 ml microcentrifuge tube the add 60 µl of Elution buffer and incubate at room temperature for 1 min then centrifuge at 8000 rpm for 1 min
- In the final step discard the mini-column and close the cap of 1.5 ml microcentrifuge containing viral RNA store at -20°C
- Next step is Master mixing and PCR
Reference: QIAamp Viral RNA Handbook
For more details: Click Here
