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Plate assay for determination of DNA concentration

Plate assay for determination of DNA concentration

A fairly accurate, rapid assay of DNA concentration can be obtained by UV visualization of samples spotted onto ethidium bromide-containing agarose plates.

1) Prepare 0.8% (w/v) agarose/ethidium bromide plates by melting 0.8g agarose in 100ml 1 x TAE buffer (50 x TAE: 2M Tris-acetate, 50mM EDTA), allowing agarose to cool to approximately 50°C and adding 10ul of a 10mg/ml ethidium bromide stock solution (final concentration was 1ug/ml). Plates can be stored for up to 1 month at 4°C in the dark.

2) Prepare standard DNA solutions of known concentrations to cover the concentration range of 10ng/ul to 200ng/ul in distilled water or TE buffer.

3) Spot standards (1ul) carefully onto the surface of a plate in duplicate followed immediately afterwards by the DNA sample of unknown concentration (1ul).

4) Allow spots to absorb for 10 to 15 minutes.

5) Visualise DNA by illumination using a shortwave UV light box.

The concentration of DNA in the unknown sample can be approximated by comparison with the standards.


Keywords: DNA, concentration

Contact Email Address: Simon.Dawson@nott.ac.uk

Submitted at 16:24 on 22/1/96 by:

Dr. Simon Dawson,
Department of Biochemistry,
University of Nottingham,
The Medical School ,
Q.M.C. ,
Clifton Boulevard ,
Nottingham,
NG7 2UH,
U.K..
Tel: +44 (0)115 9249924 ex.44787,
FAX: +44 (0)115 9422225,

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