This is a general protocol page for extracting RNA from cells.
- Endy:RNA extraction
- Extraction of RNA from tissues in English
- Extraction of RNA from tissues in French
- Gattuso:RNA extraction
Some general notes on isolating RNA.
These notes are focused on bacterial cells but please contribute to include information about plant, mammalian and other cells etc.
- Centrifugation of cells can induce a stress response in cells altering transcript levels (potentially unequally across different transcripts).
- "RNAprotect" is a reagent used to stabilize RNA content in bacterial cells grown in liquid culture.
- Works best if cells are grown in minimal media. Works worst in LB liquid culture due to complex components.
- Prepare all reagents and organize all equipment ahead of time to minimize the time between end of cell growth and cell lysis (especially if stabilization reagents are not used).
- Once cells are lysed, keep all samples on ice and use ice cold reagents to reduce RNase activity. However, note that RNases are still active at 0°C. (Do not put cells on ice prior to cell lysis otherwise you risk inducing a cold-shock response.)