Have you tried adding β-mercaptoethanol (β-ME) to the Buffer RLT before use. Add 10 µl β-ME per 1 ml Buffer RLT. This will block endogenous RNases which may be resulting in low RINs.

Mark Ian Fowler I did add the β-mercaptoethanol and I’ve tried other RNase inhibitors as well, still getting low RIN. I use RNase free everything, and have a designated RNA area that’s cleaned before use. I just can’t seem to get consistent RIN above 8. They range from 3-6. Thanks for the input, it’s a good suggestion.

Hi, Have you tried other RNA extraction kits?

Perhaps the issue lies before the extraction itself. How long does it take you to process the blood and isolate the neutrophils? This time period can impact the integrity of the RNA.

Problem solved, thanks for all the suggestions!