I've been conducting RNA extraction from 200 microliters of whole blood using the Purelink RNA Minikit according to the provided instructions. Despite several attempts, I'm consistently obtaining low yields (less than 20ng) with poor quality, indicated by a 260/280 ratio of 3+ and low 260/230 ratios. Unfortunately, I'm unable to switch to a different kit at the moment. Could someone please give me some advice on how to improve both the yield and quality of my RNA extraction?
If you follow the instruction, you should get clean RNA with expected yields (generally around > Average yields of RNA are between 2-4 µg/0.5 ml of whole blood, https://www.thermofisher.com/tr/en/home/references/ambion-tech-support/rna-isolation/tech-notes/purify-rna-directly-from-whole-blood.html#:~:text=Average%20yields%20of%20RNA%20are,RT%2DPCR%20and%20array%20analysis.). Are you using the buffy coat or the whole blood?
One little detail, are you making a fresh new Beta-mercaptoethanol before isolation (that's the stinky chemical that needs to be stored in chemical cabinets)? That has to be made fresh, can't use it after a week, a month etc.
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