RNA extraction

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This is a general protocol page for extracting RNA from samples.
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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.
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#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).
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#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 11:41, 9 October 2007

This is a general protocol page for extracting RNA from cells.

Contents

Specific Protocols

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.

  1. Centrifugation of cells can induce a stress response in cells altering transcript levels (potentially unequally across different transcripts).
  2. "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.
  3. 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).
  4. 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.

Reference

  1. Matthew B. Avison. Measuring gene expression. New York, NY: Taylor & Francis Group, 2007. isbn:0415374723. [MeasuringGeneExpression]
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