RNA extraction
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| - | This is a general protocol page for extracting RNA from | + | This is a general protocol page for extracting RNA from cells. |
==Specific Protocols== | ==Specific Protocols== | ||
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#[http://www1.qiagen.com/literature/handbooks/literature.aspx?id=1000278 "RNA''protect''"] is a reagent used to stabilize RNA content in bacterial cells grown in liquid culture. | #[http://www1.qiagen.com/literature/handbooks/literature.aspx?id=1000278 "RNA''protect''"] 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. | #*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.) | ||
==Related information== | ==Related information== | ||
Revision as of 12:41, 9 October 2007
This is a general protocol page for extracting RNA from cells.
Contents |
Specific Protocols
- Endy:RNA extraction
- Extraction of RNA from tissues in English
- Extraction of RNA from tissues in French
- Gattuso:RNA extraction
Notes
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.)
Related information
See RNA.
