RNA Quality Control: Difference between revisions
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Revision as of 06:11, 12 October 2006
Quality control of RNA
RNA Quality control This protocol will assist you in performing RNA quality control for a variet of purposes; Microarray, real-time and Northern analysis.
Checking RNA/cRNA samples on a nanodrop Spec
- Check concentration, A260/A280, A260/A230 on Nanodrop Spectrophotometer.
- Maintain the A260/A280 ratio close to 2.0 for pure RNA (ratios between 1.9 and 2.1 are acceptable).
- For Affymetrix labeling reactiosns; concentrations of cRNA are generally around 1000-1500ng/µl for a 40µl IVT reaction using 20µl of cDNA template.
Checking RNA/cRNA samples on an agilent bioanalyzer
Agilent BioAnalyzer Gel electrophoresis of the RNA is done to estimate the quality of the RNA.
RNA Image galleries
The following are examples of typical Human RNA examined on an Agilent 2100 BioAnalyser. RNA electropherograms have different profiles depending on species, tissue and starting material quality.
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Human
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Human slightly degraded
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Human very degraded
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Arabidopsis
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Human brain
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Human muscle
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Human blood
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Drosphila
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Drosphila degraded
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Arabidopsis
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Arabidopsis degraded
Explanation of electropherograms.
cRNA Image galleries
The following are examples of typical cRNA products examined on an Agilent 2100 BioAnalyser. cRNA electropherograms have different profiles depending on species and starting material quality.
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Human cRNA 10ug starting RNA
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Human cRNA 5ug starting RNA
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Human cRNA 1ug starting RNA
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Human cRNA 100ng starting RNA
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Human cRNA 10ng starting RNA
Explanation of electropherograms.
Checking RNA/cRNA Samples on a denaturing formaldehyde gel
I still need to add a formaldhyde gel protocol, anyone else got one?
