Hello, I'm working with samples stored in RNA later. Thus, I want to know do I have to do some extra step before grinding the sample into liquid nitrogen. Because when I extracted RNA from RNA later sample.The A260/230 concentration was low (between 1.4-1.7). But the A260/A280 concentration was good (Between 2-2.1). Can I use this product for RNA seq?
To improve the A260/230 ratio before grinding the samples in liquid nitrogen:
Precipitate Contaminants: Try to remove contaminants by precipitating your RNA with ethanol or isopropanol before proceeding with RNA extraction.
Use a Different RNA Extraction Method: Consider using a different RNA extraction method that is more effective at removing contaminants. Various commercial kits are available, and they may provide better RNA quality compared to standard methods.
Quality Control: After RNA extraction, assess the quality of the RNA using an Agilent Bioanalyzer, TapeStation, or a similar instrument that provides a detailed RNA profile. This will give you a clearer picture of the RNA integrity.
Regarding the use of the extracted RNA for RNA-Seq, the A260/A280 ratio is a good indicator of RNA purity, and your values (between 2-2.1) are within an acceptable range. However, for RNA-Seq, it's important to assess the RNA integrity further. The most commonly used metric for this is the RNA Integrity Number (RIN) determined by capillary electrophoresis. A RIN value above 7 is typically considered acceptable for RNA-Seq.
- RNA later do not generally impact directly the RNA concentration or quality
- RNA concentration is different from what you are mentioning.
- "A260/230 concentration" is not concentrations
- 260/230 are ratio which indicate towards the quality of DNA or RNA.
- For better estimate of the quality and quantity of RNA, try to use bioanalyzer and/or qubit like methods.
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