I am going to extract RNA from influenza virus infected cells using TRIZOL. So, I wonder whether culture media will have any interference in RNA extraction and subsequent downstream application.
Hi Prem
It is generally recommended that the cells to be used should wash with PBS before performing any experiment. PBS is just a isotonic buffer solution and it will only help you to get rid of cell debris and medium based contaminants without decreasing your RNA purity once you remove. Trizol indeed blocks RNase activity, however you should use PBS to extract high quality RNA. I also suggest you to perform DNase treatment especially working with viral RNA if your protocol is suitable for it.
Good luck
Probably if you do not remove culture media you could have protein contamination in your RNA extracts and therefore downstream interferences.
I never washed the humas or mife cells to remove excess of cell culture media.and it always worked. As far as i know The trizol inhibits all RNAse presrent in the sample. As I never worked with virus I have no idea if they can precipitewhen speeding.
Hi Prem
It is generally recommended that the cells to be used should wash with PBS before performing any experiment. PBS is just a isotonic buffer solution and it will only help you to get rid of cell debris and medium based contaminants without decreasing your RNA purity once you remove. Trizol indeed blocks RNase activity, however you should use PBS to extract high quality RNA. I also suggest you to perform DNase treatment especially working with viral RNA if your protocol is suitable for it.
Good luck
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