I want to extract RNA from human breast milk. How should I store the milk sample in RNAlater?
And after RNA extraction how can I store the RNA?
Dear Anwesha
Usually, Milk as such is not preserved. You need to separate the milk components and preserve it i.e. Somatic cells, Milk fat.
So, if your study is to evaluate expression of genes from somatic cells present in milk (somatic cells are supposed to be not present in milk, but in inflammatory state it increase in milk), you should separate somatic cells from the milk and then preserve these in RNAlater or extract RNA immediately.
Dear Anwesha,
I agree with Jitendra Kumar Shandilya .
You should use a protocol similar like this one (under cooled conditions):
-> Pipet 700µl milk and 700µl PBS(1X) in a sterile eppendorf tube (1,5ml)
-> Centrifuge for 10min at 1500g
-> Remove the supernatans with a pipet
-> Remove the leftover cream on the walls of the eppendorf tube with a sterile swab, make sure not to touch the pellet
-> Add 1ml PBS(1X) to the pellet and vortex to resuspend the pellet
-> Centrifuge for 10min at 1500g
-> Remove the supernatans with a pipet
-> Remove the leftover cream on the walls of the eppendorf tube with a sterile swab, make sure not to touch the pellet
After this you can start extraction and ultimately freeze your extract at -80°C until PCR.
Also check https://www.hindawi.com/journals/bmri/2015/475935/
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