Can I store the freshly collected tissue sample in trizol instead of RNA later buffer? if Yes, How long can i store? will it affect on quality/quantity of the RNA sample?
Which is better among RNA later and Trizol?
Can Trizol will be a substitution for RNA later?
Hello Indumathi A N Swamy,
RNAs preserved in TRIzol and RNAlater have similar RNA quality. However, for tissues, RNAlater is more convenient at the site of sample collection, while TRIzol method would be much easier at homogenizing freshly collected tissues.
For both these methods you may proceed as follows.
For RNAlater:
1. Excise the tissue sample within 10 minutes after death. Rinse in PBS and blot dry briefly. Cut it into slices less than 5 mm thick. Perform this step as quickly as possible and proceed to the next step.
2. Completely submerge the tissue pieces in the collection vessel containing RNAlater. At least 10 volumes of the reagent (or approximately 10 uL reagent per 1 mg of tissue) will be required.
3. Incubate the tissue in the reagent. With RNAlater, tissue can be stored at room temperature for up to 1 week, at 4ºC for up to 1 month and at -80ºC for a long time. RNA extraction may be done at a later date using TRIzol.
For TRIzol:
1. Excise the tissue sample within 10 minutes after death. Rinse in PBS and blot dry briefly. Cut the tissues into small pieces to ensure that the tissue is completely immersed in TRIzol reagent. Transfer tissue pieces in 1 mL TRIzol reagent per 10~25 mg tissue. The tissue volume should not exceed 10% of the TRIzol reagent volume.
2. Homogenize the tissue in TRIzol on ice using a tissue homogenizer.
3. The lysate can be stored at 4°C for days, -20°C for a year or at -80°C indefinitely.
Can TRIzol be a substitution for RNA later?
Yes, unless and until the collected tissue sample is completely homogenized in TRIzol.
RNA later is a preservative which stabilizes cellular RNA without the need to freeze samples. This solution preserves intact RNA by precipitating out RNases into an aqueous sulfate salt solution.
On the other hand, TRIzol can only inactivate RNases with which it is in direct contact. Therefore, tissue samples are not safe from RNA degradation until it is completely homogenized.
Best.
Malcolm Nobre Thank you for your detailed clarification on my query. Your Valuable comments are great help for my research.
You may keep tissue samples using Trizol (also known as TRI reagent), particularly when you want to preserve the integrity of your RNA. Storing tissue samples effectively helps you do just that.
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