Hello everyone,
I have been using the QIAamp RNA Blood Mini Kit to extract RNA from human whole blood. While I am mostly following the provided protocol, I made a couple of minor adjustments:
Despite adhering to the protocol otherwise, I am encountering consistently low RNA yields. Out of 11 attempts, only once did I achieve a concentration of 133 ng/µL, while the others were below 20 ng/µL as measured with a NanoDrop. I need a significantly higher and consistent RNA yield for downstream applications.
I have also tried the standard 20-minute lysis incubation, but this did not improve the yield.
Could these modifications be the root of the issue? Are there other potential adjustments or troubleshooting steps I could try to increase RNA yield? Any insights or advice would be greatly appreciated.
Thank you in advance!
I think if you are getting consistent yields then you might need to look more at input than the protocol. How far below your target yield are you? There are ways to increase concentration but eluting in smaller volume or concentrating the sample.
One tip:
For column elution I like to load the water (or elution buffer) and let it sit on the column for 1-2 minutes then spin. Then I will reload the eluate to the column, let sit 1-2 minutes and spin again, this can help with yield. or you elute in 1/2 the volume and elute twice rather than reloading.
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