Hello,
I've been trying to isolate RNA from hiPSC derived neuron culture (70 days old) with GenElute™Mammalian Total RNA Miniprep Kit but I could not obtain a good yield even though I had a good amount of neurons in the culture. I isolated from a single well of 6-well plate which was not fully confluent yet the networks were extensive and dense. I am sure of my homogenization step since there were not any particles.
Did you experience something unusual while isolating RNA from dense neuron cultures and do you know a way to recover RNA from the column if the sample clogged it?
Thanks in advance
One of the ways to recover RNA from a clogged column is to keep re-eluting 2 to three times. My advice will be to heat the column at 70 degrees, and keep eluting. Of cos you lose some volume, but ultimately I hope the RNA concentration that you gain from re-eluting will be higher. Another way is to do RNA precipitation overnight to increase RNA concentration.
Thank you so much for your contribution! I tried 10 minutes and 30 minutes incubation but it didn''t help:(. o you think it is too much?
Hi, my way of doing it is incubating it for 5 minutes at 70 degress celsius with the elution buffer. Usually it works well. incubating it too long at 70 degrees is going to reduce yield as evaporation will cause the volume to become too little
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