StainsAll

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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				Stains All Method

			Direct staining of DNA-fragments after electrophoresis can be useful if the results should be defined quantitatively. No UV-table and special camera is needed, a normal scanner will do the job. The quantification runs via a dilution series of a special quantitative marker

		Dilution of the Marker The marker “QS200” from GenSura, available from Carl Roth (#L508) is diluted 10 + 90, this gives 60 ng in 6 ul. Further dilution should be made down to 1 ng in 6 ul

	The Dye Stains All is available from Sigma. The order number is # E9379-1G

April 28, 2023


	The Sensitivity (per band) Double strands in native gels: 5 - 10 ng. Single strands in denatured gels: 10 - 20 ng


	The Method

Procedure	Recipe	Time (10% gel)	Time (15%, HyRes)
Washing	H₂O Bidest	10 min	15 min
Staining	16 mg Stains All, 100 ml N-Methylpyrrolidon, 100 ml H₂O. Final volume: 200 ml	1-2x 15 min*	1-2x 30 min*
Destaining / Preserving	10%Ethanol, 10 % Glycerin	20 min	20 min
Light-Destaining / Scan	Destain in normal room-light for 1.5 h. Scan in transmission mode, adjust contrast. Air-dry the gel over-night.	1.5 h	1.5 h


	The Result, run native


			*Sometimes double staining is necessary

		The Result, run denatured


	EDC Electrophoresis Development & Consulting, Vor dem Kreuzberg 17, 72070 Tübingen (Germany)